KINDAI UNIVERSITY


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NAGAI Kouhei

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FacultyDepartment of Genetic Engineering / Graduate School of Biology-Oriented Science and Technology
PositionAssociate Professor
Degree
Commentator Guidehttps://www.kindai.ac.jp/meikan/368-nagai-kouhei.html
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Last Updated :2020/09/30

Education and Career

Education

  •   1996 04  - 1999 03 , Kyoto University, Fuculty of Agriculture
  •   1999 04  - 2001 03 , Graduate School of Kyoto University, Fuculty of Agriculture
  •   2001 04  - 2004 03 , Graduate School of Kyoto University, Fuculty of Agriculture

Academic & Professional Experience

  •   2016 04 ,  - 現在, Associated professor, BOST, Kindai University
  •   2012 07 ,  - 2016 03 , Wakayama Industry promotion fundation
  •   2012 04 ,  - 2015 03 , Lecturer, BOST, Kindai University
  •   2009 09 ,  - 2012 03 , Assistant professor, St. Mariannna University of Medicine
  •   2005 04 ,  - 2009 08 , Post Doctoral Fellow, Wakayama Industry promotion fundation

Research Activities

Research Areas

  • Life sciences, Food sciences
  • Life sciences, Allergies and connective tissue disease
  • Other, Other, Laboratory medicine

Research Interests

  • Lifestyle-related diseases, Metabolic Syndrome, Functional food, animal science, post-translational modification, autoimmunity, proteomics

Published Papers

  • Signs of biological activities of 28,000-year-old mammoth nuclei in mouse oocytes visualized by live-cell imaging, Kazuo Yamagata, Kouhei Nagai, Hiroshi Miyamoto, Masayuki Anzai, Hiromi Kato, Kei Miyamoto, Satoshi Kurosaka, Rika Azuma, Igor I. Kolodeznikov, Albert V. Protopopov, Valerii V. Plotnikov, Hisato Kobayashi, Ryouka Kawahara-Miki, Tomohiro Kono, Masao Uchida, Yasuyuki Shibata, Tetsuya Handa, Hiroshi Kimura, Yoshihiko Hosoi, Tasuku Mitani, Kazuya Matsumoto, Akira Iritani, Scientific Reports, Scientific Reports, 9, 4050, Mar. 2019 , Refereed
  • Ume polyphenol: prebiotic effects on diet-induced obesity mice and growth-promoting mechanism for bifidobacteria., Masahiro Kumeno, Azusa Ito, Natsumi Ohigashi, Yuki Yoshihara, Toshio Suzuki, Kouhei Nagai, Hisashi Ashida, Mem. Faculity. B.O.S.T. Kindai University, Mem. Faculity. B.O.S.T. Kindai University, 42, 1 - 14, Nov. 2018 , Refereed
  • Ubiquitin-proteasome system modulates zygotic genome activation in early mouse embryos and influences full-term development., Higuchi C, Shimizu N, Shin SW, Morita K, Nagai K, Anzai M, Kato H, Mitani T, Yamagata K, Hosoi Y, Miyamoto K, Matsumoto K, J Reprod Dev., J Reprod Dev., 64(1), 65 - 74, Dec. 2017 , Refereed
  • Altered acetylation of proteins in patients with rheumatoid arthritis, revealed by acetyl-proteomics, Arito M, Nagai K, Ooka S, Sato T, Takakuwa Y, Kurokawa MS, Okamoto K, Suematsu N, Kato T, Arthritis Research & Therapy, Arthritis Research & Therapy, 33(6), 877 - 886, 2015 , Refereed
  • Large-scale proteomic analysis of Japanese Black cattle : Part III. Identification of protein biomarker candidates for assessment of carcass traits and meat quality characteristics, Ikegami H, Nagai K, Matsuhashi T, Kobayash N, Takemoto A, Yoshihiro T, Inoue E, Higuchi T, Morita K, Uchibori S, Amano T, Y Taguchi, Kato H, A Iritani, Matsumoto K, Japanese Society of Animal Science, Japanese Society of Animal Science, 86, 141 - 152, 2015 , Refereed
  • Roles of serum fibrinogen alpha chain-derived peptides in Alzheimer's disease, Miwa Noguchi, Toshiyuki Sato, Kouhei Nagai, Itaru Utagawa, Itsuku Suzuki, Mitsumi Arito, Nobuko Iizuka, Naoya Suematsu, Kazuki Okamoto, Tomohiro Kato, Noboru Yamaguchi, Manae S. Kurokawa, INTERNATIONAL JOURNAL OF GERIATRIC PSYCHIATRY, INTERNATIONAL JOURNAL OF GERIATRIC PSYCHIATRY, 29(8), 808 - 818, Aug. 2014 , Refereed
    Summary:Objective: To find a blood biomarker and disease-related peptides in Alzheimer's disease (AD), we comprehensively detected serum peptides. Methods: Ion intensity of serum peptides from 62 AD patients and 82 control subjects was measured by mass spectrometry. Results: A total of 157 peptides were detected from 30 AD patients and 30 healthy control (HC) subjects. Sixty out of the 157 peptide profiles discriminated between the AD and HC groups. Sixteen out of the 60 peptides were identified, 10 out of which were fragments of a fibrinogen alpha chain (FIBA). Among the 10 peptides, four and six peptides were derived from fibrinopeptide A (FPA, A alpha 1-16) and the C-terminal region of the alpha C-domain (alpha CDC, A alpha 557-610), respectively. The profile of 10 FIBA-derived peptides combined with age discriminated between the two groups with an area under the receiver operating characteristic curve (AUROC) of 0.940. Validation of this model using a testing set of 32 AD patients and 19 HC subjects showed an AUROC of 0.717, sensitivity of 65.6%, and specificity of 73.7% by a cutoff value of 0.56420. Another value, 0.04029, showed sensitivity of 96.9%, suggesting that subjects with values less than 0.04029 rarely possess AD. FPA and alpha CDC showed increased ion intensity in the AD group compared with the HC group (p < 0.05). Conclusions: The profile of 10 FIBA-derived peptides combined with age would be a candidate biomarker for AD, which facilitates screening of the disease. The significant release of FPA and alpha CDC may be involved in the aberrant coagulation that leads to vascular damage in AD. Copyright (C) 2013 John Wiley & Sons, Ltd.
  • Protein profiles of peripheral blood mononuclear cells as a candidate biomarker for Behcet's disease, T. Yoshioka, M. S. Kurokawa, T. Sato, K. Nagai, N. Iizuka, M. Arito, Y. Takakuwa, H. Nakano, S. Ooka, N. Suematsu, K. Okamoto, K. Yudoh, H. Nakamura, N. Suzuki, S. Ozaki, T. Kato, CLINICAL AND EXPERIMENTAL RHEUMATOLOGY, CLINICAL AND EXPERIMENTAL RHEUMATOLOGY, 32(4), S9 - S19, Jul. 2014 , Refereed
    Summary:Objective. To investigate the pathophysiology of Behcet's disease (BD) and find biomarkers for the disease, we analysed protein profiles of peripheral blood mononuclear cells (PBMCs). Methods. Proteins, extracted from PBMCs, were comprehensively analysed in 16 patients with BD, 16 patients with rheumatoid arthritis (RA), 12 patients with Crohn's disease (CD), and 16 healthy control subjects (HC) by 2-dimensional differential gel electrophoresis (2D-DIGE). Differently expressed proteins were identified by mass spectrometry. Results. 563 protein spots were detected. We completely discriminated between the BD and HC groups, between the BD and RA groups, and between the BD and CD groups by multivariate analysis of intensity of 23, 35, and I spots, respectively. The spots contributing to the differences included proteins related to cytoskeleton, transcription! translation, T cell activation, bone turnover, regulating apoptosis, and microbial infection. Intensity of 3 spots (tyrosine-protein phosphatase non-receptor type 4, threonine synthase-like 2, and beta-actin) provided area under the receiver operating characteristic curves (AUROC) of 0.889 for discrimination between the BD group and the non-BD groups. Informatively, intensity of the above I spot completely discriminated the CD group from the other groups (AUROC 1.000). This spot, identified as beta-actin, had different pI from the above beta-actin-spot probably due to different post-translational modification. Conclusion. PBMC protein profiles, especially the profile of the 3 spots, would be candidate biomarkers for BD. The latter beta-actin subtype would be useful for discriminating inflammatory bowel diseases from BD and other diseases. The identified proteins may play important roles in the pathophysiology of BD.
  • Serum peptides, represented by complement 3f des-arginine, are useful for prediction of the response to pegylated interferon-α plus ribavirin in patients with chronic hepatitis C, Yohei Noguchi, Manae S. Kurokawa, Chiaki Okuse, Nobuyuki Matsumoto, Kouhei Nagai, Toshiyuki Sato, Mitsumi Arito, Naoya Suematsu, Kazuki Okamoto, Michihiro Suzuki, Fumio Itoh, Tomohiro Kato, Hepatology Research, Hepatology Research, 43(7), 743 - 756, Jul. 2013 , Refereed
    Summary:Aim: Biomarkers predicting sustained virological response (SVR) to pegylated interferon-α plus ribavirin (PEG IFN-α/RBV) were investigated. Methods: Peptides in pretreatment sera from 107 patients with hepatitis C virus (HCV) genotype 1 were comprehensively analyzed by mass spectrometry. Ion intensity of the peptides was used to generate discriminant models between the responders who achieved SVR (R) and the non-responders (NR) to PEG IFN-α/RBV. Results: In total, 107 peptides were detected in a training set (n = 23). A discriminant model using a peptide, complement 3f des-arginine (C3f-dR), showed sensitivity of 35% and specificity of 94% for SVR prediction in a testing set (n = 68). In all the R and NR (n = 96), an area under the receiver-operator curve (AUROC) of 0.64 in the C3f-dR model was increased to 0.78 by addition of platelet (PLT) counts (C3f-dR/PLT model). Another model using the 107 peptides (AUROC, 0.77) also showed higher AUROC (0.79) by addition of hemoglobin (Hb), body mass index (BMI) and age (107P/Hb/BMI/Age model). The sensitivity and specificity of the C3f-dR/PLT model were 59% and 88%, and those of the 107P/Hb/BMI/Age model were 70% and 92%, respectively. The C3f-dR/PLT model showed high AUROC (0.82), similar to that of interleukin-28B rs8099917 genotype analysis (0.86) in the 45 tested patients. Prediction by the combination of the C3f-dR/PLT model, the 107P/Hb/BMI/Age model and the rs8099917 genotype analysis was accurate in 44 out of the 45 patients (AUROC, 0.95). Conclusion: Serum peptides, especially C3f-dR, would be useful predictors for SVR to PEG IFN-α/RBV. The complements may be involved in the HCV elimination. © 2012 The Japan Society of Hepatology.
  • Protein profiles of peripheral blood mononuclear cells are useful for differential diagnosis of ulcerative colitis and Crohn's disease, Moriaki Hatsugai, Manae S. Kurokawa, Takefumi Kouro, Kohei Nagai, Mitsumi Arito, Kayo Masuko, Naoya Suematsu, Kazuki Okamoto, Fumio Itoh, Tomohiro Kato, JOURNAL OF GASTROENTEROLOGY, JOURNAL OF GASTROENTEROLOGY, 45(5), 488 - 500, May 2010 , Refereed
    Summary:Effective biomarkers for discrimination between ulcerative colitis (UC) and Crohn's disease (CD) have not been established yet. In this study, we analyzed protein profiles of peripheral blood mononuclear cells (PBMCs) of the patients to find such a biomarker. Peripheral blood mononuclear cell proteins from 17 UC patients, 13 CD patients, and 17 healthy controls were separated by two-dimensional gel electrophoresis. The intensities of individual protein spots were subjected to discriminant analysis of UC and CD using the SIMCA-P+program. We found that 547 protein spots were commonly detected among the UC, CD, and healthy groups. Orthogonal partial least squares-discriminant analysis using 276 protein spots clearly discriminated the UC patients from the CD patients (R (2) 0.994; Q (2) 0.462). A similar analysis using a further selected 58 protein spots showed higher performance for discrimination of the diseases (R (2) 0.948; Q (2) 0.566). Eleven out of the 58 protein spots were successfully identified; these were functionally related to inflammation, oxidation/reduction, the cytoskeleton, endocytotic trafficking, and transcription. In addition, the PBMC protein profiles were useful for the prediction of disease activity in the UC and the CD patients, and they were also useful for predicting disease severity and responses to treatments in the UC patients. PBMC protein profiles are useful for the discrimination of UC from CD. The profiles could be a potent biomarker for the differential diagnosis of these diseases. Further investigation of the proteins which contributed to the discrimination could promote elucidation of the pathophysiology of UC and CD.
  • Seasonal changes in proteomic profiles of Japanese kelp: Saccharina japonica (laminariales, phaeophyceae)., Yotsukura N, Nagai K, Kimura H, Morimoto K, J Appl Phycol., J Appl Phycol., 21, 265 - 272, 2009 , Refereed
  • Developing an integrated database system for the large-scale proteomic analysis of Japanese Black cattle, Nagai K, Yoshihiro T, Inoue E, Ikegami H, Sono Y, Kawaji H, Kobayashi N, Matsuhashi T, Morimoto K, Nakagawa S, Iritani A, Matsumoto K, Japanese Society of Animal Science, Japanese Society of Animal Science, 79, 467 - 481, 2008 , Refereed
  • Peroxiredoxin as a functional endogenous antioxidant enzyme in pronuclei of mouse zygotes, Kohtaro Morita, Mikiko Tokoro, Yuki Hatanaka, Chika Higuchi, Haruka Ikegami, Kouhei Nagai, Masayuki Anzai, Hiromi Kato, Tasuku Mitani, Yoshitomo Taguchi, Kazuo Yamagata, Yoshihiko Hosoi, Kei Miyamoto, Kazuya Matsumoto, Journal of Reproduction and Development, Journal of Reproduction and Development, 64(2), 161 - 171, 2018 , Refereed
    Summary:Antioxidant mechanisms to adequately moderate levels of endogenous reactive oxygen species (ROS) are important for oocytes and embryos to obtain and maintain developmental competence, respectively. Immediately after fertilization, ROS levels in zygotes are elevated but the antioxidant mechanisms during the maternal-to-zygotic transition (MZT) are not well understood. First, we identified peroxiredoxin 1 (PRDX1) and PRDX2 by proteomics analysis as two of the most abundant endogenous antioxidant enzymes eliminating hydrogen peroxide (H2O2). We here report the cellular localization of hyperoxidized PRDX and its involvement in the antioxidant mechanisms of freshly fertilized oocytes. Treatment of zygotes at the pronuclear stage with H2O2 enhanced pronuclear localization of hyperoxidized PRDX in zygotes and concurrently impaired the generation of 5-hydroxymethylcytosine (5hmC) on the male genome, which is an epigenetic reprogramming event that occurs at the pronuclear stage. Thus, our results suggest that endogenous PRDX is involved in antioxidant mechanisms and epigenetic reprogramming during MZT.
  • Serum peptides as putative modulators of inflammation in psoriasis, Tetsuhiko Matsuura, Masaaki Sato, Kouhei Nagai, Toshiyuki Sato, Mitsumi Arito, Kazuki Omoteyama, Naoya Suematsu, Kazuki Okamoto, Tomohiro Kato, Yoshinao Soma, Manae S. Kurokawa, JOURNAL OF DERMATOLOGICAL SCIENCE, JOURNAL OF DERMATOLOGICAL SCIENCE, 87(1), 36 - 49, Jul. 2017 , Refereed
    Summary:Background: Psoriasis is a refractory inflammatory disease, however, its pathophysiology is still not fully understood. Objective: We tried to identify novel serum peptides associated with the pathophysiology of psoriasis. Methods: Serum peptides from 24 patients with psoriasis vulgaris (PV), 10 patients with psoriatic arthritis (PsA),14 patients with atopic dermatitis (AD), and 23 healthy control (HC) subjects were analyzed by mass spectrometry. The effects of some peptides on the secretion of humoral factors from dermal cells were investigated by cytokine arrays and ELISAs. Results: A total of 93 peptides were detected. 24, 20, 23, and 2 peptides showed at least 1.2-fold difference in ion intensity between the psoriasis (PV + PsA) and HC groups, between the PV + PsA and AD groups, between the PV and PsA groups, and between patients with severe-to-moderate PV (n = 6) and those with mild PV (n = 18), respectively (p < 0.05). 13 out of 27 peptides that showed at least 1.5-fold ion intensity difference in the abovementioned 4 comparisons were identified. The parent proteins of the identified peptides included a coagulation factor,, proteins involved in the maintenance of skin, and a protein relating to cytoskeleton. We focused on 2 peptides that were increased in the PV + PsA group: a fibrinogen a chain-derived peptide (1462 m/z), the unmodified form of which was fibrinopeptide A-desalanine (FPAdA), and a filaggrin (FLG)-derived peptide (1977 m/z), a modified form of FLG(2099-2118) (Q(2099)pE, Q(2115)E; FLG-pEE). FPAdA stimulation increased the secretion of GRO alpha from dermal microvascular endothelial cells (dMVECs) and decreased the secretion of lipocalin-2 from keratinocytes in comparison to FPAdA-resequenced peptide stimulation (GRO alpha, 280.9 +/- 7.3 pg/mL vs. 229.6 +/- 5.0 pg/mL, p < 0.001; lipocalin-2, 273 +/- 13 pg/mL vs. 350 +/- 10 pg/mL, p < 0.01). Interestingly, FLG-pEE stimulation decreased the secretion of GRO alpha, IL-8, and MCP-1 from dMVECs in comparison to FLG-derived control peptide stimulation (GRO alpha, 844.3 +/- 47.5 pg/mL vs. 1038.5 +/- 96.9 pg/mL, p < 0.05; IL-8, 2240.1 +/- 172.6 pg/mL vs. 3221.8 +/- 523.7 pg/mL, p < 0.05; MCP-1, 4057.8 +/- 157.2 pg/mL vs. 4619.1 +/- 213.4 pg/mL, p < 0.05). Conclusions: The results suggested that some serum peptides are involved in the pathophysiology of psoriasis, regulating the secretion of inflammatory chemokines and an antimicrobial protein. The modulation of serum peptides may be a potential therapeutic strategy for psoriasis. (C) 2017 Japanese Society for Investigative Dermatology. Published by Elsevier Ireland Ltd. All rights reserved.
  • OXIDATIVE MODIFICATION OF MYELOPEROXIDASE IN ANTI-NEUTROPHIL CYTOPLASMIC ANTIBODY (ANCA)-ASSOCIATED VASCULITIDES, Manae S. Kurokawa, Kouhei Nagai, Toshiyuki Sato, Masaaki Sato, Yukiko Takakuwa, Seido Ooka, Mitsumi Arito, Tomohiro Kato, RHEUMATOLOGY, RHEUMATOLOGY, 56, 113 - 113, Mar. 2017 , Refereed
  • Comparative proteomic analysis of neutrophils from patients with microscopic polyangiitis and granulomatosis with polyangiitis, Teisuke Uchida, Kouhei Nagai, Toshiyuki Sato, Nobuko Iizuka, Mitsumi Arito, Yukiko Takakuwa, Hiromasa Nakano, Seido Ooka, Manae S. Kurokawa, Naoya Suematsu, Kazuki Okamoto, Shoichi Ozaki, Tomohiro Kato, Journal of Proteomics, Journal of Proteomics, 91, 259 - 269, Oct. 08 2013 , Refereed
    Summary:Both microscopic polyangiitis (MPA) and granulomatosis with polyangiitis (GPA) belong to ANCA-associated vasculitis (AAV), in which neutrophils play a key role in their pathology. In this study, in order to discriminate between MPA and GPA, protein profiles of peripheral blood polymorphonuclear cells (PMNs) of 11 MPA patients and 9 GPA patients and 10 healthy controls (HC) were analyzed by 2D-DIGE. In all the 864 spots detected, intensity of 55 spots was significantly different (p< . 0.05) among the three groups by ANOVA. 31 out of the 55 spots were identified by mass spectrometry. Orthogonal partial-least-squares-discriminate analysis revealed that the abundance profile of the protein spots discriminated the AAV group from the HC group, and the MPA group from the GPA group completely. 13 protein spots were considered as biomarker candidates to distinguish between MPA and GPA. In those, spots whose intensity was higher in MPA than in GPA included actin with various p. I values, while a considerable part of spots whose intensity was higher in GPA were proteins related with the activity of neutrophils. Among the candidate proteins, ROC analysis showed that a combination of neutrophil gelatinase-associated lipocalin and a-kinase anchor protein 7 isoforms beta had a high diagnostic potential. Biological significance: In this study, protein profiles of polymorphonuclear cells (PMNs) of microscopic polyangiitis (MPA) and granulomatosis with polyangiitis (GPA) patients and healthy controls (HC) were investigated by 2D-DIGE, and MS analysis. As a result, we found that the protein profiles of PMNs were useful for distinguishing between patients (MPA and GPA) and HC, and between patients with MPA and patients with GPA. Especially, we found that the 13 protein spots that consisted of 10 proteins considerably contributed to the discrimination between MPA and GPA. This is the first to demonstrate that protein profiles of PMNs are different among MPA, GPA and healthy control. The 10 proteins we identified in this study would be new biomarkers for the diagnosis of the diseases, and may be reflect the pathology difference between MPA and GPA. © 2013 Elsevier B.V.
  • Protein Profiles Of Peripheral Blood Mononuclear Cells As a Biomarker For Behcet's Disease, Manae Kurokawa, Takuya Yoshioka, Toshiyuki Sato, Kouhei Nagai, Nobuko Iizuka, Mitsumi Arito, Yukiko Takakuwa, Hiromasa Nakano, Seido Ooka, Naoya Suematsu, Kazuki Okamoto, Hiroshi Nakamura, Noboru Suzuki, Shoichi Ozaki, Tomohiro Kato, ARTHRITIS AND RHEUMATISM, ARTHRITIS AND RHEUMATISM, 65, S811 - S812, Oct. 2013 , Refereed
  • Investigation of Proteomic Profiles of Lamina of Ecklonia kurome (Laminariales): Homology-Based Cross-Species Protein Identification and Analysis of the Post-translational Processing of Vanadium-Dependent Bromoperoxidases Using MALDI-TOF/TOF, Kouhei Nagai, Koichi Morimoto, Haruka Ikegami, Hajime Kimura, Norishige Yotsukura, MARINE BIOTECHNOLOGY, MARINE BIOTECHNOLOGY, 15(4), 487 - 498, Aug. 2013 , Refereed
    Summary:Proteomic profiles of the lamina of Ecklonia kurome Okamura, one of the Japanese dominant laminarialean kelps, were investigated by two-dimensional electrophoresis (2-DE) and MALDI-TOF/TOF. Due to the absence of E. kurome DNA or protein databases, homology-based cross-species protein identification was performed using a combination of three database-searching algorithms, Mascot peptide mass fingerprinting, Mascot MS/MS ion search, and mass spectrometry-based BLAST. Proteins were extracted from the lamina by an ethanol/phenol method and subjected to 2-DE (pI 4-7, 10 % polyacrylamide gel). More than 700 spots were detected in the 2-DE gel with CBB, and 93 spots (24 proteins) were successfully identified by MALDI-TOF/TOF and the cross-species database searching. The identified proteins mainly consisted of cytoplasmic carbohydrate metabolic enzymes, chloroplast proteins involved in photosynthesis, and haloperoxidases. Interestingly, vanadium-dependent bromoperoxidases (vBPO), which is thought to be involved in halogen uptake, synthesis of halogenated products, and detoxification of reactive oxygen species, were separated into at least 23 different spots. By comparing mass spectra, amino acid sequences predicted from tandem mass spectra and haloperoxidase activities of the vBPOs, we found that (1) at least two types of vBPOs were expressed in the lamina of E. kurome and (2) two pro-vBPOs might be activated by specific cleavage at N- and C-terminal regions.
  • Comparative Proteomic Analysis of Neutrophils Between Microscopic Polyangiitis and Granulomatosis with Polyangiitis., Teisuke Uchida, Kouhei Nagai, Toshiyuki Sato, Mitsumi Arito, Nobuko Iizuka, Manae Kurokawa, Naoya Suematsu, Kazuki Okamoto, Shoichi Ozaki, Tomohiro Kato, ARTHRITIS AND RHEUMATISM, ARTHRITIS AND RHEUMATISM, 64(10), S655 - S656, Oct. 2012 , Refereed
  • Altered posttranslational modification on U1 small nuclear ribonucleoprotein 68k in systemic autoimmune diseases detected by 2D Western blot, Kouhei Nagai, Mitsumi Arito, Yukiko Takakuwa, Seido Ooka, Toshiyuki Sato, Manae S. Kurokawa, Kazuki Okamoto, Teisuke Uchida, Naoya Suematsu, Tomohiro Kato, ELECTROPHORESIS, ELECTROPHORESIS, 33(13), 2028 - 2035, Jul. 2012 , Refereed
    Summary:Anti-ribonucleoprotein (anti-RNP) antibodies are one of the representative autoantibodies detectable in patients with systemic lupus erythematosus (SLE) and mixed connective tissue disease (MCTD). Generally, posttranslational modifications (PTMs) on autoantigens are proposed to be involved in the production of autoantibodies. In this study, we tried to detect the alteration in PTMs on a U1 small nuclear RNP 68k subunit (U1-68k), a major antigen of anti-RNP antibodies. Peripheral blood mononuclear cells (PBMCs) were obtained from patients with MCTD, SLE, and rheumatoid arthritis (RA), and from healthy donors. U1-68ks in the PBMCs were detected by 2D Western blot (WB), where extracted nuclear proteins were separated by 2DE, followed by the detection of U1-68k using WB. More than 20 PTM isoforms were detected with different molecular weights of 65.0 , 66.5, and 68.0kDa, and different pIs between 6.0 and 8.5. Importantly, the relative intensity of the spot with 66.5 kDa and pI 7.5 was significantly increased in the MCTD and SLE groups compared to the RA and healthy groups. Further, this U1-68k isoform, in particular, in its RS domain, was found to have significantly decreased phosphorylation compared to the other isoforms. The PTM alternation may be one of the steps to generate the anti-RNP antibodies.
  • Temperature stress-induced changes in the proteomic profiles of Ecklonia cava (Laminariales, Phaeophyceae), Norishige Yotsukura, Kouhei Nagai, Toshimitsu Tanaka, Hajime Kimura, Kouichi Morimoto, JOURNAL OF APPLIED PHYCOLOGY, JOURNAL OF APPLIED PHYCOLOGY, 24(2), 163 - 171, Apr. 2012 , Refereed
    Summary:Proteomic profiling on Ecklonia cava Kjellman grown under various seawater temperatures was conducted to search for biomarkers that were useful to evaluate the health of the colonies and formulate actions for the maintenance of marine forests. In the cultivated strains, protein expression was not significantly changed when the cultivation temperature was lowered from 15A degrees C (control) to 10A degrees C. On the contrary, it was markedly changed, i.e., photosynthesis-related proteins were up-regulated and metabolic enzymes were down-regulated, when the temperature was heightened to 20A degrees C. With the cultivation at 30A degrees C, 25 spots within 27 spots expressed at this temperature peculiarly could be identified and classified into ten proteins. Of the distinctive 27 spots at 30A degrees C, 20 spots were detected in the wild strains cultured at the same temperature for a brief time. It is presumed that the proteins including vanadium-dependent bromoperoxidase are heat stress-induced proteins.
  • AC13, a C-Terminal Fragment of Apolipoprotein A-I, Is a Candidate Biomarker for Microscopic Polyangiitis, Yukiko Takakuwa, Manae S. Kurokawa, Seido Ooka, Toshiyuki Sato, Kouhei Nagai, Mitsumi Arito, Naoya Suematsu, Kazuki Okamoto, Hiroko Nagafuchi, Hidehiro Yamada, Shoichi Ozaki, Tomohiro Kato, ARTHRITIS AND RHEUMATISM, ARTHRITIS AND RHEUMATISM, 63(11), 3613 - 3624, Nov. 2011 , Refereed
    Summary:Objective. Microscopic polyangiitis (MPA) is necrotizing vasculitis of unknown etiology. We analyzed the serum peptide profile of MPA to find a biomarker for this disease. Methods. Serum peptides from 33 patients with MPA, 7 with granulomatosis with polyangiitis (Wegener's), 7 with Churg-Strauss syndrome, 6 with giant cell arteritis, and 25 with systemic lupus erythematosus (SLE) were comprehensively analyzed by mass spectrometry. Peptide function on human microvascular endothelial cells (HMVECs) was examined by enzyme-linked immunosorbent assay and real-time polymerase chain reaction. Results. A total of 102 serum peptides were detected from the 78 patients. One of the peptides, peptide 1,523, showed significantly higher ion intensity in MPA (mean +/- SD 46.8 +/- 39.3 arbitrary units [AU]) than in the other systemic vasculitides (14.1 +/- 12.2 AU) (P < 0.05) or in SLE (17.0 +/- 12.1 AU) (P < 0.05). In MPA, peptide 1,523 showed significantly higher ion intensity before treatment than 1 week (P < 0.05) and 6 weeks (P < 0.05) after the initiation of treatment. Peptide 1,523 was identified as 13 C-terminal amino acid residues of apolipoprotein A-I (Apo A-I) and was designated "AC13." Validation of AC13 ion intensity using another MPA cohort (n = 14) similarly showed significantly higher ion intensity (90.1 +/- 167.9 AU) compared to 14 patients with rheumatoid arthritis (8.6 +/- 5.4 AU) (P < 0.01) and 14 healthy subjects (11.8 +/- 6.1 AU) (P < 0.01). Serum concentrations of Apo A-I and hig-density lipoprotein cholesterol were down-regulated in MPA before treatment and returned to their normal ranges 6 weeks after the initiation of treatment (both P < 0.01). Stimulation of HMVECs with AC13 significantly up-regulated secretion of interleukin-6 (IL-6) (P < 0.05) and IL-8 (P < 0.01). Conclusion. AC13, a candidate biomarker for MPA, may be useful for monitoring disease activity and may exacerbate vascular inflammation through upregulation of proinflammatory cytokines.
  • Proteomic analyses of aortic wall in patients with abdominal aortic aneurysm, T. Ando, K. Nagai, M. Chikada, K. Okamoto, M. S. Kurokawa, T. Kobayashi, T. Kato, H. Makuuchi, JOURNAL OF CARDIOVASCULAR SURGERY, JOURNAL OF CARDIOVASCULAR SURGERY, 52(4), 545 - 555, Aug. 2011 , Refereed
    Summary:Aim. The mechanisms underlying the formation of abdominal aortic aneurysms have yet to be fully clarified. To identify key proteins generally involved in aneurysmal formation, proteomic profiles were compared between aneurysmal and non-aneurysmal regions of aortic walls from patients with abdominal aortic aneurysm. Methods. Aortic wall specimens were obtained from three patients with abdominal aortic aneurysm. Protein profiles of aortic wall samples including vascular media and adventitia were compared between aneurysmal and non-aneurysmal regions in each patient using two-dimensional fluorescence difference gel electrophoresis (2D-DIGE). Protein spots expressed differently between the two regions were identified by tandem mass spectrometry and verified by immunohistochemical investigations. Results. Image analysis of 2D-DIGE gels revealed 22 proteins spots expressed differently between aneurysmal and non-aneurysmal regions in all three patients. Among these, five protein spots that were up-regulated in the AA regions were successfully identified as complement component C4, fragments of the fibrinogen alpha or beta subunits, and actin. immunohistochemical studies showed massive deposition of fibrin/fibrinogen or its fragments in the media, and complement C1q component, the molecule starting the classical complement pathway, in all three layers of the aneurysmal region. Conclusion. Our proteomic and subsequent immunohistochemical studies revealed significant fibrinogenesis and fibrinolysis in the media, and activation of the classical complement pathway in all three layers of the aneurysmal region. These data promote understanding of mechanisms behind the formation of abdominal aortic aneurysms.
  • Arthritogenicity of annexin VII revealed by phosphoproteomics of rheumatoid synoviocytes, Kosuke Matsuo, Mitsumi Arito, Koji Noyori, Hiroshi Nakamura, Manae S. Kurokawa, Kayo Masuko, Kazuki Okamoto, Kouhei Nagai, Naoya Suematsu, Kazuo Yudoh, Moroe Beppu, Tomoyuki Saito, Tomohiro Kato, ANNALS OF THE RHEUMATIC DISEASES, ANNALS OF THE RHEUMATIC DISEASES, 70(8), 1489 - 1495, Aug. 2011 , Refereed
    Summary:Objective To identify novel proteins involved in the pathogenesis of rheumatoid arthritis (RA) and to characterise the identified proteins based on pathogenic and therapeutic aspects. Methods The authors applied differential phosphoproteomic analysis to articular synoviocytes between RA and osteoarthritis (OA) to identify proteins differently phosphorylated between RA and OA. Focusing on annexin VII (Anx7), one of the highly phosphorylated proteins in RA, the authors prepared Anx7-transgenic C57BL/6 (Anx7-Tg-B6) mice to evaluate their susceptibility to collagen-induced arthritis (CIA). In addition, the authors examined the effect of anti-Anx7 antibodies (Abs) on CIA and serum levels of cytokines in wild-type DBA/1J mice, which are known to be susceptible to CIA, and in Anx7-Tg-B6 mice. In vitro, the authors examined the effect of the Anx7 knockdown by small interfering RNA on the secretion of cytokines in rheumatoid synoviocytes and the human synovial sarcoma cell line SW982. Results The Anx7 transgene altered the CIA-resistant B6 mice to CIA-susceptible ones. The Abs treatment suppressed CIA even in the wild-type DBA/1J mice. The serum levels of cytokines including interleukin 6 (IL-6) and TNF alpha were not altered by the Abs treatment in vivo. On the other hand, the knockdown of Anx7 by small interfering RNA caused downregulation of IL-8 secretion in vitro. Conclusions These results indicate that Anx7 participates in the pathogenesis of RA partly through the secretion of IL-8. The study data have demonstrated the pathogenic roles and therapeutic significance of Anx7 in RA for the first time.
  • Identification and characterization of an oocyte factor required for development of porcine nuclear transfer embryos, Kei Miyamoto, Kouhei Nagai, Naoya Kitamura, Tomoaki Nishikawa, Haruka Ikegami, Nguyen T. Binh, Satoshi Tsukamoto, Mai Matsumoto, Tomoyuki Tsukiyama, Naojiro Minami, Masayasu Yamada, Hiroyoshi Ariga, Masashi Miyake, Tatsuo Kawarasaki, Kazuya Matsumoto, Hiroshi Imai, PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 108(17), 7040 - 7045, Apr. 2011 , Refereed
    Summary:Nuclear reprogramming of differentiated cells can be induced by oocyte factors. Despite numerous attempts, these factors and mechanisms responsible for successful reprogramming remain elusive. Here, we identify one such factor, necessary for the development of nuclear transfer embryos, using porcine oocyte extracts in which some reprogramming events are recapitulated. After incubating somatic nuclei in oocyte extracts from the metaphase II stage, the oocyte proteins that were specifically and abundantly incorporated into the nuclei were identified by mass spectrometry. Among 25 identified proteins, we especially focused on a multifunctional protein, DJ-1. DJ-1 is present at a high concentration in oocytes from the germinal vesicle stage until embryos at the four-cell stage. Inhibition of DJ-1 function compromises the development of nuclear transfer embryos but not that of fertilized embryos. Microarray analysis of nuclear transfer embryos in which DJ-1 function is inhibited shows perturbed expression of P53 pathway components. In addition, embryonic arrest of nuclear transfer embryos injected with anti-DJ-1 antibody is rescued by P53 inhibition. We conclude that DJ-1 is an oocyte factor that is required for development of nuclear transfer embryos. This study presents a means for identifying natural reprogramming factors in mammalian oocytes and a unique insight into the mechanisms underlying reprogramming by nuclear transfer.
  • Micro-Sieving: Isolation of Whole Glomeruli from a Single Renal Needle Biopsy Sample, Kenichiro Koitabashi, Kazuki Okamoto, Mitsumi Arirto, Toshiyuki Sato, Kouhei Nagai, Manae S. Kurokawa, Naoya Suematsu, Takashi Yasuda, Kenjiro Kimura, Tomohiro Kato, NEPHRON CLINICAL PRACTICE, NEPHRON CLINICAL PRACTICE, 117(3), C225 - C229, 2011 , Refereed
    Summary:Renal biopsy samples are important not only for the diagnosis of glomerulonephritis, but also for the investigation of its pathogenesis. However, it remains difficult to biochemically analyze proteins extracted solely from the glomeruli of needle biopsy samples, since the samples contain various components like renal tubules and connective tissue. Even a recent micro-dissection method, recovering the glomeruli in the sliced sections of the biopsy samples, has not fully solved the difficulty because the amount of obtainable proteins by this method is not usually enough for protein analysis. To overcome this problem, we established a simple but reliable method to isolate whole glomeruli from needle biopsy samples. By this method, termed 'micro-sieving', we were able to isolate on average more than 50 glomeruli from a single needle biopsy sample in an hour. The amount of the extracted glomerular proteins was on average 23 mu g per biopsy sample. As a representative use of this method, we were able to obtain a glomerular protein profile by fluorescent 2-dimensional electrophoresis for each of the tested patients with glomerulonephritis. 'Micro-sieving' can be used widely as a fundamental technique to analyze glomeruli in renal needle biopsy samples. Copyright (C) 2010 S. Karger AG, Basel
  • Actinidain-hydrolyzed Type I Collagen Reveals a Crucial Amino Acid Sequence in Fibril Formation, Saori Kunii, Koichi Morimoto, Kouhei Nagai, Takuya Saito, Kenji Sato, Ben'ichiro Tonomura, JOURNAL OF BIOLOGICAL CHEMISTRY, JOURNAL OF BIOLOGICAL CHEMISTRY, 285(23), 17465 - 17470, Jun. 2010 , Refereed
    Summary:We investigated the ability of type I collagen telopeptides to bind neighboring collagen molecules, which is thought to be the initial event in fibrillogenesis. Limited hydrolysis by actinidain protease produced monomeric collagen, which consisted almost entirely of alpha 1 and alpha 2 chains. As seen with ultrahigh resolution scanning electron microscopy, actinidain-hydrolyzed collagen exhibited unique self-assembly, as if at an intermediate stage, and formed a novel suprastructure characterized by poor fibrillogenesis. Then, the N- and C-terminal sequences of chicken type I collagen hydrolyzed by actinidain or pepsin were determined by Edman degradation and de novo sequence analysis with matrix-assisted laser desorption ionization-tandem time-of-flight mass spectrometry, respectively. In the C-telopeptide region of the alpha 1 chain, pepsin cleaved between Asp(1035) and Phe(1036), and actinidain between Gly(1032) and Gly(1033). Thus, the actinidain-hydrolyzed alpha 1 chain is shorter at the C terminus by three residues, Gly(1033), Phe(1034), and Asp(1035). In the alpha 2 chain, both proteases cleaved between Glu(1030) and Val(1031). We demonstrated that a synthetic nonapeptide mimicking the alpha 1 C-terminal sequence including GFD weakly inhibited the self-assembly of pepsin-hydrolyzed collagen, whereas it remarkably accelerated that of actinidain-hydrolyzed collagen. We conclude that the specific GFD sequence of the C-telopeptide of the alpha 1 chain plays a crucial role in stipulating collagen suprastructure and in subsequent fibril formation.
  • Identification of autoantigens specific for systemic lupus erythematosus with central nervous system involvement, N. Iizuka, K. Okamoto, R. Matsushita, M. Kimura, K. Nagai, M. Arito, M. S. Kurokawa, K. Masuko, N. Suematsu, S. Hirohata, T. Kato, LUPUS, LUPUS, 19(6), 717 - 726, May 2010 , Refereed
    Summary:Using proteomic analysis, we identified candidate autoantigens specific for central nervous system (CNS) involvement in systemic lupus erythematosus (SLE). Proteins, extracted from cultured human neuroblastoma cells, were separated both by SDS-PAGE (1-DE) and two-dimensional electrophoresis (2-DE), and transferred to membranes. Western blot analysis was performed using serum samples from 30 SLE patients with CNS involvement (CNS-Lupus) and from 30 SLE patients without CNS involvement (non-CNS-SLE). The detected autoantigens were identified using MALDI-TOF/TOF MS. On the 1-DE Western blot, we detected 32 antigenic bands in the serum samples from the CNS-Lupus patients. Among them, four bands were detected significantly more frequently in the CNS-Lupus patients than in the non-CNS-SLE patients. Three bands were detected in four or more of the CNS-Lupus patients but in only one or none of the non-CNS-SLE patients. We thus selected these seven bands for the next investigations. Next, we detected protein spots corresponding to the selected seven bands by 2-DE Western blot and identified four proteins. They are peroxiredoxin-4, ubiquitin carboxyl-terminal hydrolase isozyme L1, splicing factor arginine/serine-rich 3, and histone H2A type 1. These four candidate autoantigens for the anti-neuronal cell antibodies would be a useful marker for CNS-Lupus. Lupus (2010) 19, 717-726.
  • Comprehensive analysis of short peptides in sera from patients with IgA nephropathy, Nagayuki Kaneshiro, Yang Xiang, Kouhei Nagai, Manae S. Kurokawa, Kazuki Okamoto, Mitsumi Arito, Kayo Masuko, Kazuo Yudoh, Takashi Yasuda, Naoya Suematsu, Kenjiro Kimura, Tomohiro Kato, RAPID COMMUNICATIONS IN MASS SPECTROMETRY, RAPID COMMUNICATIONS IN MASS SPECTROMETRY, 23(23), 3720 - 3728, Dec. 2009 , Refereed
    Summary:We analyzed serum short peptides comprehensively to know whether they were useful to characterize IgA nephropathy (IgAN). Serum samples from 26 patients with untreated IgAN and 25 healthy donors were tested. Short peptides with molecular weights of similar to 7kDa, purified from the serum samples by magnetic-beads-based weak cation exchange, were detected by mass spectrometry. Then the peptide peaks detected were subjected to the multivariate data analysis by SIMCA-P+(R) containing principal component analysis (PCA) and orthogonal partial-least-squares-discriminate analysis (OPLS-DA). A total of 92 peptide peaks were detected in the tested serum samples. The OPLS-DA analysis revealed that the profile of all the peptide peak intensities discriminated the IgAN group and the healthy group completely with a high R2 value (0.919) and a high Q2 value (0.861). Further, the profile of only five peptide peaks was found to discriminate the two groups. By tandem mass spectrometry and database searching, three of the five peptides which increased in the IgAN group were identified as fragments of fibrinogen alpha chain, and the two peptides which increased in the healthy group were identified as fragments of complement C3f and kininogen-1 light chain. Taken together, the profile of the serum short peptides would be useful to discriminate IgAN and healthy conditions. Further, the five peptides may be candidate serum markers for IgAN and may be related to pathogenesis of IgA. Copyright (c) 2009 John Wiley & Sons, Ltd.
  • Proteomic Analysis of the Mouse Ovary in Response to Two Gonadotropins, Follicle-Stimulating Hormone and Luteinizing Hormone, Manabu Satoh, Mikiko Tokoro, Haruka Ikegami, Kouhei Nagai, Youhei Sono, Seung-Wook Shin, Satoshi Nishikawa, Kazuhiro Saeki, Yoshihiko Hosoi, Akira Iritani, Aisaku Fukuda, Yoshiharu Morimoto, Kazuya Matsumoto, JOURNAL OF REPRODUCTION AND DEVELOPMENT, JOURNAL OF REPRODUCTION AND DEVELOPMENT, 55(3), 316 - 326, Jun. 2009 , Refereed
    Summary:Functional and structural changes in the mammalian ovary are coordinately regulated by the pituitary glycoprotein hormones, follicle-stimulating hormone (FSH) and luteinizing hormone (LH), leading to follicular development, ovulation and transformation of follicles into corpus lutea. To investigate protein profiles during these processes of the mouse ovarian cycle, we applied combined methods (two-dimensional gel electrophoresis [2-DE] for separation and visualization of proteins plus matrix laser desorption/ionization time-of-flight mass spectrometry [MALDI-TOF/MS] analysis for protein identification) for comparative proteomic analysis using immature mice at 3 weeks of age. Protein profiles were obtained from proteins extracted from intact ovaries that had been collected from pregnant mare serum gonadotropin (PMSG)/human chorionic gonadotropin (hCG)-primed immature mice at 0 (no PMSG), 24 and 48 h post PMSG, as well as at 10 and 20 h post hCG. The results showed that 1028 common protein spots were found in representative gels that had been separated in the 3 to 11 pH range and the 15-200 kDa range, 253 protein spots (24.6%) of which were differentially expressed (p<0.05) during the mouse ovarian cycle. Of these 253 protein spots, 99 were identified by MALDI-TOF/MS. This comparative proteomic approach to identifying proteins that were potentially involved in the complex process of the ovarian cycle could contribute to our understanding of the molecular basis of functional and structural changes in the ovary in response to gonadotropins. Furthermore, the interesting ovarian proteins identified in this study may eventually serve as diagnostic biomarker candidates of ovarian function.
  • Proteomic analysis of the rat cerebellar flocculus during vestibular compensation, Masahiko Fukasawa, Kazuki Okamoto, Manabu Nakamura, Koshi Mikami, Sonoko Shimada, Yasuhiko Tanaka, Kouhei Nagai, Mitsumi Arito, Manae S. Kurokawa, Kayo Masuko, Naoya Suematsu, Izumi Koizuka, Tomohiro Kato, JOURNAL OF VESTIBULAR RESEARCH-EQUILIBRIUM & ORIENTATION, JOURNAL OF VESTIBULAR RESEARCH-EQUILIBRIUM & ORIENTATION, 19(3-4), 83 - 94, 2009 , Refereed
    Summary:Unilateral labyrinthectomy (UL) in rats is used as a human vertigo model. In this model, spontaneous nystagmus and dysequilibrium caused by UL are ameliorated within 48-72 hours. The amelioration, termed vestibular compensation (VC), is long lasting. Although cerebellar flocculi have been reported to be involved in VC, the molecular mechanisms behind VC are unknown. In this study, we used 2D-DIGE to detect protein changes in flocculi during acute (48 hours) and chronic (1 week) stages of VC. We found 99 out of 967 protein spots that showed significant changes in their intensities. Of the 99 spots, 45 spots (ipsilateral side, 15; contralateral side, 30) changed unilaterally during the acute stage, whereas 46 spots (ipsilateral side, 21; contralateral side, 25) changed unilaterally during the chronic stage. Thus, the acute compensation mechanism is more complicated in the contralateral flocculus than in the ipsilateral flocculus. Using MALDI-TOF MS, we identified 10 proteins out of the 12 protein spots. Of these, 3 proteins involved in synaptic transmission, neuronal filament formation and vesicular transport, respectively, demonstrated VC generation.
  • Protein extraction for 2-DE from the lamina of Ecklonia kurome (laminariales): Recalcitrant tissue containing high levels of viscous polysaccharides, Kouhei Nagai, Norishige Yotsukura, Harulka Ikegami, Hajime Kimura, Koichi Morimoto, ELECTROPHORESIS, ELECTROPHORESIS, 29(3), 672 - 681, Feb. 2008 , Refereed
    Summary:Extraction of proteins from the tissues of laminarialean algae, i.e. kelp, is difficult due to high levels of nonprotein interfering compounds, mainly viscous polysaccharides. To establish proteomic analysis of kelp species, an ethanol/phenol extraction method was developed and compared to other popular methods. Proteins were extracted with phenol from crude protein powder, obtained by homogenizing the kelp tissues in ice-cold ethanol. The ethanol/phenol method produced high-quality proteins of the highest purity from the lamina of Ecklonia kurome, one of the Japanese dominant laminarialean algae. This method gave well-resolved 1-D SDS-PAGE or 2-DE images with low background and the highest number of bands or spots. In particular, proteins with neutral to basic pI's were efficiently extracted. Furthermore, 27 spots on the 2-DE gel were extensively identified by MALDI-TOF/TOF analysis. To the best of our knowledge, this is the first report of a protocol for protein extraction from kelp tissues that gives satisfactory 2-D protein profiles. It is expected that the protocol can be applied to other algae tissues or other recalcitrant plant tissues containing high levels of nonprotein interfering compounds.
  • A novel nacre protein N19 in the pearl oyster Pinctada fucata, Masato Yano, Kouhei Nagai, Koichi Morimoto, Hiroshi Miyamoto, BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 362(1), 158 - 163, Oct. 2007 , Refereed
    Summary:A novel 19 kDa protein, which was named N19, was isolated from the nacreous layer of the pearl oyster Pinetada fucata. N19 is one of predominant proteins found in the water-insoluble fraction of the nacreous layer. MALDI-TOF/TOF analysis indicated that the three trypsin-digested peptides (791.45, 824.42, and 1118.65 m/z) corresponded to the amino acid sequences predicted from a cDNA isolated from a mantle cDNA library of P. fucata. Northern blot analysis revealed that the N19 mRNA was a little more abundant in the pallial region than the edge region, in the mantle. In CaCO3 precipitation assay, the recombinant N19 protein inhibited the crystallization of CaCO3. These results indicate that N19 is localized in the nacre and plays a negative regulatory role in calcification in the pearl oyster. (C) 2007 Elsevier Inc. All rights reserved.
  • Tyrosinase localization in mollusc shells, Kouhei Nagai, Masato Yano, Koichi Morimoto, Hiroshi Miyamoto, COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY B-BIOCHEMISTRY & MOLECULAR BIOLOGY, COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY B-BIOCHEMISTRY & MOLECULAR BIOLOGY, 146(2), 207 - 214, Feb. 2007 , Refereed
    Summary:In molluscan shellfish, pigmentation is frequently observed in the calcified shell, but the molecular basis of this process is not understood. Here, we report two tyrosinase proteins (Pfty1 and Pfty2) found in the prismatic shell layer of the pearl oyster Pinctada fucata; this layer is recognized as the pigmented region in P. fucata. The protein sequences were deduced from the corresponding cDNAs and confirmed by MALDI-TOF/TOF analysis. The sequences suggest that both tyrosinases have two copper-binding sites in similar N-terminal domains that are homologous to tyrosinases of cephalopods and hemocyanins of gastropods. In turn, this suggests that bivalve tyrosinases are evolved from a common ancestral copper-binding protein in the mollusc. Pfty1 and Pfty2 were specifically expressed in the mantle, and their expression in the mantle is different from each other, suggesting that these tyrosinases have distinctive roles in melanogenesis in shells. (c) 2006 Elsevier Inc. All rights reserved.
  • Shematrin: A family of glycine-rich structural proteins in the shell of the pearl oyster Pinctada fucata, M Yano, K Nagai, K Morimoto, H Miyamoto, COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY B-BIOCHEMISTRY & MOLECULAR BIOLOGY, COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY B-BIOCHEMISTRY & MOLECULAR BIOLOGY, 144(2), 254 - 262, Jun. 2006 , Refereed
    Summary:Random sequencing of molecules from a cDNA library constructed from mantle mRNA of the pearl oyster Pinctada fucata was used to obtain information on organic matrix proteins in the shell. In the determined sequences, we identified 7 distinct cDNAs encoding similar glycine-rich domains. Complete sequence analysis of these cDNAs showed that the predicted sequences of the proteins, which we named shematrins, possessed similar domains comprising repeat sequences of two or more glycines, followed by a hydrophobic amino acid. In addition, in shematrin-1, -2 and -3, a repeat domain designated as XG(n)X (where X is a hydrophobic amino acid) was conserved. It is of further note that all the shematrin proteins have RKKKY, RRKKY or RRRKY as their C-terminal sequence. According to northern blot analysis, all shematrins are exclusively expressed in the mantle, and particularly in the edge region of the mantle-, furthermore, peptide fragments similar to shematrin-1 and -2 were detected in the prismatic layer of shells by MALDI-TOF/TOF MS analysis. These findings suggest that many of shematrins are synthesized in the mantle edge and secreted into the prismatic layer of the shell, where the protein family is thought to provide a framework for calcification. (c) 2006 Elsevier Inc. All rights reserved.
  • Insights into the reaction mechanism of the coagulation of soy protein isolates induced by subtillsin Carlsberg, K Nagai, K Inouye, JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY, JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY, 52(15), 4921 - 4927, Jul. 2004 , Refereed
    Summary:The reaction mechanism of the coagulation of soy protein isolates (SPIs) induced by subtilisin Carlsberg was investigated. Formation of the coagula was monitored by measuring the turbidity (OD660) of the SPI solution, which decreased at the initial stage (phase 1 or digestion phase) of the reaction, and then increased (phase 2 or coagulation phase) and finally reached the plateau level. The velocity of the coagulation increased with increasing enzyme concentration. The coagulation was inhibited dramatically by adding a serine protease inhibitor (phenylmethanesulfonyl fluoride, PMSF) when the turbidity reached the minimum value. This indicates that the SPI digests participating in the coagulation are produced mainly in phase 2; in other words, production of the coagulating fragments and their coagulation occur simultaneously in phase 2. Structural changes of SPI during proteolysis were measured by observing fluorescence changes of aromatic amino acids of SPI and an externally added hydrophobic probe. It was suggested that the hydrophilic surface areas of SPIs might be cleaved preferentially in phase 1, and that the hydrophobic inner areas might be cleaved in phase 2 with extensive decomposition of the 3-D structure of SPI proteins. The fragments formed in phase 2 are considered to coagulate through hydrophobic interactions.
  • Coagulation of soy protein isolates induced by subtilisin Carlsberg, K Inouye, K Nagai, T Takita, JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY, JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY, 50(5), 1237 - 1242, Feb. 2002 , Refereed
    Summary:The coagulation of soy protein isolates (SPI) induced by subtilisin Carlsberg was studied. The proteins were digested to fragments of 16 kDa or less in the early stage of the reaction, followed by coagulation. The time-course of the coagulation measured by turbidity was separated into three phases. The turbidity decreased from the initial level observed at time zero to the minimum level (OD1) at time T1 (15-20 min) in the first phase. Then, it increased drastically to reach the maximum (OD2) at time T2 (60-70 min) in the second phase, which was followed by a slight decrease in the third phase. The coagulation was terminated at T2, where 30-35% of the weight of the SPI proteins was in coagula. Proteins in the coagula were degraded slowly in the prolonged incubation, and the protein content in the coagula was finally 15-20% of the weight. The time-course of the turbidity agreed well with that of the weight of the precipitates formed, indicating that the turbidity reflects the progress of the coagulation. The turbidity change (OD1 to OD2) from the start to the end of the coagulation increased proportionally to the SPI concentration (4.9-11 mg/mL), although the time (T1 to T2) needed for the coagulation was independent of the concentration. The growth of the coagula is promoted by increasing the SPI concentration and is rate-limiting in the coagulation.

Conference Activities & Talks

  • Comprehensive analysis of the effects of high-fat diet on protein abundance in the mouse liver using SWATH acquisition method., S. Yamawaki, N. Misawa, T. Nishibata, R. Nagamine, T. Awaji, Y. Oosedo, A. Sakaue, K. Kishida, K. Nagai, JPROS2019,   2019 07 26
  • Comprehensive analysis of post-translational modifications on myeloperoxidase from patients with ANCA associated vasculitis., R. Sasaki, K. Nagai, T. Nishibata, M. Sato, T. Sato, M. S. Kurokawa, T. Kato, JPROS2019,   2019 07 26
  • Comprehensive analysis of post-translational modifications on the proteins from a 28,000-year-old woolly mammoth, T. Nishibata, K. Nagai, K. Yamagata, H. Miyamoto, M. Anzai, H. Kato, K. Miyamoto, R. Azuma, I. I. Kolodeznikov, A. V. Protopopov, V. V. Plotnikov, Y. Hosoi, T. Mitani, K. Matsumoto, A. Iritani, JPROS2019,   2019 07 26
  • Proteomic analysis of muscle and bone marrow tissues obtained from a 28,000-year-old woolly mammoth., K. Nagai, H. Miyamoto, M. Anzai, R. Azuma, T. Nishibata, K. Yamagata, H. Kato, K. Miyamoto, I. I. Kolodeznikov, A. V. Protopopov, V. V. Plotnikov, Y. Hosoi, T. Mitani, K. Matsumoto, A. Iritani, JPROS2019,   2019 07 25
  • Comprehensive quantification of liver proteins in the high-fat induced-obese mouse using a SWATH-MS analysis, Tomoya Nishibata, Sayaka Yamawaki, Siomi Oota, Tomoki Awaji, Yuki Oosedo, Ayaka Sakaue, Kunihiro Kishida, Kouhei Nagai,   2018 09 25 , 招待有り
  • Discovery of biomarker candidate proteins for predicting beef carcass traits and meat quality characteristics by time-course proteomic analysis of serum in Japanese Black steers using SWATH-MS and Lasso regression analysis., H. Ikegami, T. Matsuhashi, K. Ochi, H. Sano, K. Nagai, K. Obayashi, H. Kato, S. Sakaguchi, T. Yoshihiro, K. Masumoto, MSP2018,   2018 05 16
  • Assessment of reproducibility and quantitative performance of SWATH-mass spectrometry of bovine serum proteins., K. Makino, T. Nishibata, S. Yamawaki, H. Ikegami, T. Matsuhashi, K. Nagai, K. Matsumoto, MSP2018,   2018 05 16
  • Comprehensive quantification of liver proteins in the high-fat induced-obese mouse using a SWATH acquisition method – Toward elucidation of molecular mechanism of functional food substances-, S. Oota, ○S. Yamawaki, T. Nishibata, T. Awaji, Y. Oosedo, A. Sakaue, K. Kishida, K. Nagai, MSP2018,   2018 05 15
  • New extraction approach of non-collagenous proteins from bone tissue by mild-degradation of enzyme, Saori Kunii, Kouhei Nagai, Koichi Morimoto, ECTS 2017,   2017 05 14
  • Oxidative modification of myeloperoxidase in anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitides, Manae S. Kurokawa, Kouhei Nagai, Toshiyuki Sato, Masaaki Sato, Yukiko Takakuwa, Seido Ooka, Mitsumi Arito, Tomohiro Kato, The 18th International Vasculitis and ANCA Workshop,   2017 03
  • Prediction of beef carcass traits using changes in amount of serum proteins during fattening period, H ikegami, T Matsuhashi, K Nagai, M Morimoto, T Tsukamoto, S Yamaguchi, C Higuchi, K Morita, S Sakamoto, K Matsumoto, JPROS2016,   2016 07 29
  • なし, Kouhei Nagai,   2016 04 21
  • なし, Kouhei Nagai, Nobuhei Ryu, Koji Yoshida, Tsuyoshi Itani, Mizuki Higashiyama,   2016 03 29
  • 第37回日本分子生物学会,   2014 08
  • Protein profilrs of peripheral blood mononuclear cells as a biomarker for bechet's disease., Takuya Yoshioka, Manae S. Kurokawa, Toshiyuki Sato, Kouhei Nagai, Nobuko Iizuka, Mitsumi Arito, Yukiko Takakuwa, Hiromasa Nakano, Seido Ooka, Naoya Suematsu, Kazuki Okamoto, ACR/ARHP annual meeting 2013,   2013 10
  • Comparative proteomic analysis of neutrophils from patients with microscopic polyangiitis and granulomatosis with polyangiitis., Teisuke Uchida, Kouhei Nagai, Toshiyuki Sato, Nobuko Iizuka, Mitsumi Arito, Yukiko Takakuwa, Hirosama Nakano, Seido Ooka, Manae Kurokawa, Naoya Suematsu, Kazuki Okamoto, Shoichi Ozaki, Tomohiro Kato, ACR/ARHP annual meeting 2013,   2013 10
  • Proteomic analysis of muscle tissue for discovery of proteins related to accumulation of intramuscular fat in beef cattle, Atsushi TAKEMOTO, Kouhei NAGAI, Haruka IKEGAMI, Natsumi SHIMIZU, Kohtaro MORITA, Chika HIGUCHI, Eiji KOBAYASHI, Kazuya MATSUMOTO, Animal Science Day 2013,   2013 09
  • Proteomic analysis of bovine serum: Discovery of biomarker proteins evaluating carcass and meat quality traits in Japanese Black beef cattle., Kouhei NAGAI, Haruka IKEGAMI, Atsushi TAKEMOTO, Natsumi SHIMIZU, Kohtaro MORITA, Chika HIGUCHI, Eiji KOBAYASHI, Kazuya MATSUMOTO, Animal Science Day 2013,   2013 09
  • Comprehensive Analysis of Aberrantly Glycosylated Proteins in Rheumatoid Arthritis, Toshiyuki Sato, Mitsumi Arito, Manae S. Kurokawa, Yukiko Takakuwa, Seido Ooka, Kouhei Nagai, Hiroshi Nakamura, Nobuko Iizuka, Naoya Suematsu, Kazuki Okamoto, Tomohiro Kato, HUPO2013,   2013 09
  • Protein Profiles of Peripheral Blood Mononuclear Cells as a Biomarker for Behcet’s Disease, Takuya Yoshioka, Manae S. Kurokawa, Toshiyuki Sato, Kouhei Nagai, Nobuko Iizuka, Mitsumi Arito, Yukiko Takakuwa, Hiromasa Nakano, Seido Ooka, Naoya Suematsu, Kazuki Okamoto, HUPO2013,   2013 09
  • Identification of Proteins Related to Accumulation of Intramuscular Fat in Japanese Black by Proteomic Analysis, Atsushi Takemoto, Kouhei Nagai, Haruka Ikegami, Natsumi Shimizu, Kohtaro Morita, Chika Higuchi, Eiji Kobayashi, Kazuya Matsumoto,   2013 09
  • Biomarker Discovery from Low Abundance Proteins in Bovine Serum; Proteomics for Developing Beef Production Systems, Haruka Ikegami, Kouhei Nagai, Atsushi Takemoto, Natsumi Shimizu, Kohtaro Morita, Chika Higuchi, Tamako Matsuhashi, Naohiko Kobayashi, Kazuya Matsumoto,   2013 09
  • Comparative Proteomic Analysis of Neutrophils from Patients with Microscopic Polyangiitis and Granulomatosis with Polyangiitis, Teisuke Uchida, Kouhei Nagai, Toshiyuki Sato, Nobuko Iizuka, Mitsumi Arito, Yukiko Takakuwa, Hirosama Nakano, Seido Ooka, Manae Kurokawa, Naoya Suematsu, Kazuki Okamoto, Shoichi Ozaki, Tomohiro Kato,   2013 09
  • Serum Peptides, Represented by Complement 3f Des-Arginine, Are Useful for Prediction of the Response to Pegylated Interferon-α Plus Ribavirin in Patients with Chronic Hepatitis C, Yohei Noguchi, Manae S. Kurokawa, Chiaki Okuse, Nobuyuki Matsumoto, Kouhei Nagai, Toshiyuki Sato, Mitsumi Arito, Naoya Suematsu, Kazuki Okamoto, Michihiro Suzuki, Fumio Itoh, Tomohiro Kato, HUPO2013,   2013 09
  • Altered acetylation of proteins in patients with rheumatoid arthritis, revealed by acetyl-proteomics., Arito M, Nagai K, Ooka S, Sato T, Takakuwa Y, Kurokawa MS, Okamoto Suematsu N, Arito M, Nagai K, Ooka S, Sato T, Takakuwa Y, Kurokawa MS, Okamoto Suematsu N, Kato T,   2012 12
  • Comparative proteomic analysis of neutrophils between microscopic polyangiitis and granulomatosis with polyandiiti, Yoshioka T, Kurokawa MS, Sato T, Iiduka N, Arito M, Nagai K, Suematsu N, Okamoto K, Kato T, ACR/ARHP annual meeting 2012,   2012 10
  • Comprehensive analysis of protein expression in peripheral blood mononuclear cells from patients with Bechet's Disease., Kouhei NagaiYoshioka T, Kurokawa MS, Sato T, Iiduka N, Arito M, Nagai K, Suematsu N, Okamoto K, Kato T, ACR/ARHP annual meeting 2012,   2012 10
  • Altered acetylation of proteins in patients with rheumatoid arthritis, revealed by acetyl-proteomics., Arito M, Nagai K, Ooka S, Takakuwa Y, Kurokawa MS, Sato T, Iiduka N, Okamoto K, Suematsu N, Kato T, HUPO2012,   2012 09
  • Screening of glycoproteins with altered glycans in rheumatoid arthritis., Sato T, Ooka S, Takakuwa Y, Nagai K, Arito M, Iizuka N, Kurokawa MS, Okamoto K, Suematsu N, Kato T, HUPO2012,   2012 09
  • Comparative proteomic analysis of neutrophils from patients with microscopic polyangiitis and granulomatosis with polyangiitis., Uchida T, Nagai K, Takakuwa Y, Ooka S, Arito M, Satoh T, Kurokawa MS, Okamoto K, Suematsu N, Kato T, HUPO2012,   2012 09
  • Comprehensive analysis of protein expression in peripheral blood mononuclear cells from patients with Bechet's disease., Kurokawa MS, Yoshioka T, Sato T, Iiduka N, Arito M, Nagai K, Suematsu N, Okamoto K, Kato T, HUPO2012,   2012 09
  • Adenosine Deaminase identified as a citrullination-dependent autoantigen in rheumatoid arthritis, by proteomic surveillance with in vitro citrullination., Kato T, Mitsui H, Arito M, Sato T, Suematsu N, Okamoto K, Kurokawa MS, Yudo K, Nagai K, Beppu M, HUPO2012,   2012 09
  • Oxidative modification on myeloperoxidase in patients with Anineutrophil cytoplasmic antibody (ANCA)-associated Vascullitides., Nagai K, Uchida T, Takakuwa Y, Ooka S, Arito M, Satoh T, Kurokawa MS, Okamoto K, Suematsu N, Kato T, HUPO2012,   2012 09
  • Comprehensive analysis of serum peptides in patients with Alzheimer's Disease., Noguchi M, Kurokawa MS, Utagawa I, Nagai K, Sato T, Arito M, Suematsu N, Okamoto K, Yamaguchi N, Kato T,   2012 09
  • 日本プロテオーム学会 日 2012年大会,   2012 07
  • Comparative proteomic analysis of neutrophils from patients with microscopic polyangiitis and granulomatosis with polyangiitis., Teisuke Uchida, Kouhei Nagai, Yukiko Takakuwa,   2012 04

Misc

  • Comprehensive quantification of liver proteins in the high-fat induced-obese mouse using a SWATH acquisition method, Shiomi Oota, Tomoya Nishibata, Sayaka Yamawaki, Tomoki Awaji, Yuki Oosedo, Ayaka Sakaue, Kunihiro Kishida, Kouhei Nagai, Mem. Faculity. B.O.S.T. Kindai University, 42, 15, 32,   2018 11 , Refereed
  • 柿ポリフェノールの脂質代謝改善効果とそのメカニズム解明, 鈴木利雄, 鈴木利雄, 大東夏海, 伊藤あずさ, 吉原侑希, 米野雅大, 永井宏平, 米谷俊, 石島智子, 阿部啓子, 岡田晋治, 森口仁文, 芦田久, 日本食品科学工学会大会講演集, 65th, 115,   2018 08 22 , http://jglobal.jst.go.jp/public/201802217429682479
  • 畜産領域へのリキッドバイオプシーの展開 2:牛の枝肉成績を肥育中に予測する血清バイオマーカータンパク質の探索, 松橋珠子, 池上春香, 越智浩介, 大林賢伍, 佐野文美, 森隆史, 本廣多胤, 奥野智美, 神谷拓磨, 永井宏平, 宮本圭, 吉廣卓哉, 坂口慎一, 松本和也, 日本畜産学会大会講演要旨, 124th, 184,   2018 03 28 , https://jglobal.jst.go.jp/detail?JGLOBAL_ID=201802243958807831
  • 畜産領域へのリキッドバイオプシーの展開 1:牛の血清中タンパク質の同時定量解析方法の確立, 越智浩介, 池上春香, 松橋珠子, 大林賢伍, 永井宏平, 坂口慎一, 松本和也, 日本畜産学会大会講演要旨, 124th, 183,   2018 03 28 , https://jglobal.jst.go.jp/detail?JGLOBAL_ID=201802247513376644
  • 好中球Myeloperoxidaseの等電点を変化させる酸化修飾の同定と定量, 橋本茜, 尾上裕太郎, 浄弘由紀子, 西野芽玖, 黒川真奈絵, 加藤智啓, 永井宏平, 日本生化学会大会(Web), 90th, ROMBUNNO.3LBA‐024 (WEB ONLY), 024],   2017 12 , http://jglobal.jst.go.jp/public/201702225874461448
  • 尋常性乾癬の病態に関与する血清ペプチドの同定, 佐藤政秋, 松浦哲彦, 永井宏平, 佐藤利行, 有戸光美, 表山和樹, 末松直也, 加藤智啓, 相馬良直, 黒川真奈絵, 日本生化学会大会(Web), 90th, ROMBUNNO.3P‐1029 (WEB ONLY), 1029],   2017 12 , http://jglobal.jst.go.jp/public/201702238964699777
  • 受精後のユビキチン・プロテアソーム系による母性タンパク質の分解はマウス初期胚発生に重要である, 樋口智香, 守田昂太郎, 山口壮輝, 松橋珠子, 永井宏平, 安齋政幸, 安齋政幸, 山縣一夫, 細井美彦, 宮本圭, 松本和也, 日本卵子学会誌, 2, 1, S48, S48,   2017 04 01 , http://jglobal.jst.go.jp/public/201702240011137048
  • マウス初期胚発生過程におけるH3R2me2sの役割, 守田昂太郎, 樋口智香, 山口壮輝, 松橋珠子, 松橋珠子, 永井宏平, 安齋政幸, 安齋政幸, 加藤博巳, 加藤博巳, 山縣一夫, 細井美彦, 宮本圭, 松本和也, 日本卵子学会誌, 2, 1, S47, S47,   2017 04 01 , http://jglobal.jst.go.jp/public/201702274754974238
  • 乾癬性関節炎における血清ペプチドプロファイルの解析, 黒川真奈絵, 佐藤政秋, 永井宏平, 佐藤利行, 有戸光美, 表山和樹, 加藤智啓, 日本リウマチ学会総会・学術集会プログラム・抄録集, 61st, 804, 804,   2017 03 18 , http://jglobal.jst.go.jp/public/201702251135347763
  • Study on preservation of animal tissue provided from zoos : Toward knowledge sharing with the zoos and aquariums, Masayuki Anzai, Rika Azuma, Moriyoshi Kubo, Noriyuki Nonoue, Kazutoshi Takami, Minoru Miyashita, Hitoshi Murai, Kouhei Nagai, Akari Obashi, Takuya Orisugi, Yoshihiko Hosoi, Memoirs of Institute of Advanced Technology, Kindai University, 22, 25, 34,   2017 03 , Refereed
  • 黒毛和種去勢牛の枝肉形質を生体評価するバイオマーカー候補タンパク質の解析, 池上春香, 松橋珠子, 永井宏平, 塚口智将, 内堀翔, 樋口智香, 守田昂太郎, 小林直彦, 松本和也, 松本和也, 日本畜産学会大会講演要旨, 121st, 243,   2016 03 27 , http://jglobal.jst.go.jp/public/201602220631479224
  • ANCA関連血管炎における好中球ミエロペルオキシダーゼの酸化修飾, 永井宏平, 佐藤利行, 佐藤政秋, 高桑由希子, 大岡正道, 表山和樹, 有戸光美, 黒川真奈絵, 加藤智啓, 日本リウマチ学会総会・学術集会プログラム・抄録集, 60th, 533, 533,   2016 03 18 , http://jglobal.jst.go.jp/public/201602209191573696
  • なし, Kouhei Nagai, Inflammation&Immunity, 24, 2, 95, 99,   2016 03 , 招待有り
  • Prediction of beef carcass traits using changes in amount of serum proteins during fattening period, Ikegami Haruka, Matsumoto Kazuya, Matsuhashi Tamako, Nagai Kouhei, Morimoto Manabu, Tsukaguchi Tomomasa, Yamaguchi Soki, Higuchi Chika, Morita Kotaro, Sakaguchi Shin-ichi, Abstracts for Annual Meeting of Japanese Proteomics Society, 2016, 0, 130, 130,   2016 , http://ci.nii.ac.jp/naid/130007529297
  • 黒毛和種牛の枝肉形質を予測するバイオマーカー候補タンパク質の肥育期間中の動態, 池上春香, 松橋珠子, 永井宏平, 塚口智将, 内堀翔, 樋口智香, 守田昂太郎, 小林直彦, 松本和也, 松本和也, 日本畜産学会大会講演要旨, 120th, 89,   2015 09 11 , http://jglobal.jst.go.jp/public/201502201571862170
  • アルツハイマー病におけるフィブリノゲンα鎖由来ペプチドの役割, 野口美和, 佐藤利行, 永井宏平, 宇田川至, 鈴木慈, 有戸光美, 飯塚進子, 末松直也, 岡本一起, 加藤智啓, 山口登, 黒川(鈴木)真奈絵, 聖マリアンナ医科大学雑誌, 43, 1, 15, 16,   2015 05 , http://jglobal.jst.go.jp/public/201502204362745602
  • 全身性自己免疫疾患(膠原病)の新規診断マーカーの開発を目的とした自己抗原U1‐68Kの疾患階異的低リン酸化体の簡便な定量法の検討, NAGAI KOHEI, SAITO KAZUKI, ARITO MITSUMI, KUROKAWA MANAE, KATO TOMOHIRO, Mem Fac Biol Oriented Sci Technol Kinki Univ, 35, 7, 16,   2015 03 31 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=201502242123983524
  • ANCA関連血管炎における好中球ミエロペルオキシダーゼの酸化修飾, 内田貞輔, 永井宏平, 佐藤利行, 大岡正道, 有戸光美, 尾崎承一, 黒川真奈絵, 加藤智啓, 日本リウマチ学会総会・学術集会プログラム・抄録集, 59th, 518, 518,   2015 03 20 , http://jglobal.jst.go.jp/public/201502256653043396
  • 黒毛和種牛の脂肪交雑度を予測する血清マイクロRNAの探索, 池上 春香, 丹羽 尚人, 永井 宏平, 塚口 智将, 樋口 智香, 守田 昂太郎, 松橋 珠子, 小林 直彦, 松本 和也, 日本畜産学会大会講演要旨集, 119回, 185, 185,   2015 03
  • A transient inhibition of proteasome pathway entails a delay in the onset of ZGA and impairs full term development of mice, HIGUCHI Chika, SHIMIZU Natsumi, MORITA Kohtaro, UCHIBORI Sho, TSUKAGUCHI Tomomasa, NAGAI Kohei, ANZAI Masayuki, YAMAGATA Kazuo, HOSOI Yoshihiko, MIYAMOTO Kei, MATSUMOTO Kazuya, The Journal of Reproduction and Development Supplement, 108, 0, P, 12-P-12,   2015 , http://ci.nii.ac.jp/naid/130005492035
  • Involvement of Peroxiredoxin (Prdx) in reducing hydrogen peroxide in pronuclei of mouse zygotes., MORITA Kohtaro, TOKORO Mikiko, HIGUCHI Chika, UCHIBORI Sho, TSUKAGUCHI Tomomasa, NAGAI Kohei, ANZAI Masayuki, YAMAGATA Kazuo, HOSOI Yoshihiko, MIYAMOTO Kei, MATSUMOTO Kazuya, The Journal of Reproduction and Development Supplement, 108, 0, P, 15-P-15,   2015 , http://ci.nii.ac.jp/naid/130005492029
  • Availability of genetic resource using somatic cell nuclei derived from muscle tissues by expression of phosphorylated H2A.X, AZUMA Rika, MIYASHITA Minoru, NAGAI Kouhei, NAKAGAWA Takao, KAJIMOTO Mizuki, INOUE Tatsuya, HOSOI Yoshihiko, ANZAI Masayuki, The Journal of Reproduction and Development Supplement, 108, 0, P, 81-P-81,   2015 , http://ci.nii.ac.jp/naid/130005492100
  • Time-course proteomic analysis of cattle serum during fattening: Identification of protein biomarker candidates for assessment of beef marbling characteristics, Ikegami Haruka, Nagai Kouhei, Matsuhashi Tamako, Kobayashi Naohiko, Higuchi Chika, Morita Koutaro, Uchibori Sho, Tsukaguchi Tomomasa, Kato Hiromi, Abstracts for Annual Meeting of Japanese Proteomics Society, 2015, 0, 234, 234,   2015 , http://ci.nii.ac.jp/naid/130007498791
  • 関節リウマチにおける糖蛋白質の糖鎖構造の変異, 佐藤利行, 有戸光美, 黒川真奈絵, 高桑由希子, 大岡正道, 永井宏平, 中村洋, 飯塚進子, 岡本一起, 加藤智啓, 日本リウマチ学会総会・学術集会プログラム・抄録集, 58th, 674, 674,   2014 03 24 , http://jglobal.jst.go.jp/public/201402273708688183
  • 黒毛和種牛の脂肪交雑度を予測するタンパク質バイオマーカーの探索, 池上 春香, 松橋 珠子, 赤尾 大樹, 武本 淳史, 永井 宏平, 内堀 翔, 小林 直彦, 松本 和也, 日本畜産学会大会講演要旨集, 118回, 151, 151,   2014 03
  • マウス始原生殖細胞におけるGSEタンパク質の発現解析, MORITA KOTARO, HATANAKA YUKI, SHIMIZU NATSUMI, NISHIHARA TAKUSHI, TAKEMOTO ATSUSHI, HIGUCHI CHIKA, UCHIHORI SHO, AMANO TOMOKO, NAGAI KOHEI, KISHIGAMI SATOSHI, KATO HIROKI, MITANI TASUKU, HOSOI YOSHIHIKO, MATSUMOTO KAZUYA, J Reprod Dev, 59, Suppl Japanese Issue, J114,   2013 08 20 , 10.14882/jrds.106.0.P-26.0, http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=201302273473633087
  • 血清ペプチドC3f-dRは、C型慢性肝炎患者におけるPeg-Interferon-α+Ribavirin併用療法の応答予測に有用である, 松本 伸行, 野口 陽平, 黒川 真奈絵, 永井 宏平, 有戸 光美, 佐藤 利行, 末松 直也, 岡本 一起, 奥瀬 千晃, 鈴木 通博, 伊東 文生, 加藤 智啓, 日本臨床分子医学会学術総会プログラム・抄録集, 50回, 62, 62,   2013 04
  • ANCA関連血管炎患者における好中球ミエロペルオキシダーゼの酸化修飾, 永井宏平, 内田貞輔, 高桑由希子, 大岡正道, 有戸光美, 佐藤利行, 黒川真奈絵, 岡本一起, 加藤智啓, 日本リウマチ学会総会・学術集会・国際リウマチシンポジウムプログラム・抄録集, 57th-22nd, 527, 527,   2013 03 19 , http://jglobal.jst.go.jp/public/201302272760346629
  • 関節リウマチにおける糖蛋白質の糖鎖構造の変異, 佐藤利行, 高桑由希子, 大岡正道, 永井宏平, 有戸光美, 飯塚進子, 黒川真奈絵, 岡本一起, 加藤智啓, 日本リウマチ学会総会・学術集会・国際リウマチシンポジウムプログラム・抄録集, 57th-22nd, 557, 557,   2013 03 19 , http://jglobal.jst.go.jp/public/201302286798186413
  • ベーチェット病患者の末梢血単核球に発現している蛋白質の網羅的解析, 黒川真奈絵, 吉岡拓也, 佐藤利行, 永井宏平, 飯塚進子, 有戸光美, 高桑由希子, 中野弘雅, 大岡正道, 岡本一起, 中村洋, 鈴木登, 尾崎承一, 加藤智啓, 日本リウマチ学会総会・学術集会・国際リウマチシンポジウムプログラム・抄録集, 57th-22nd, 302,   2013 03 19 , http://jglobal.jst.go.jp/public/201302245965487325
  • アポリポ蛋白A‐IのC末端断片であるAC13の顕微鏡的多発血管炎のバイオマーカーとしての可能性, 高桑由希子, 黒川真奈絵, 大岡正道, 永井宏平, 有戸光美, 佐藤利行, 末松直也, 岡本一起, 永渕裕子, 山田秀裕, 尾崎承一, 加藤智啓, 聖マリアンナ医科大学雑誌, 40, 4, 304, 304,   2013 03 , http://jglobal.jst.go.jp/public/201302243350283336
  • マウス始原生殖細胞で発現するGSEタンパク質の能動的DNA脱メチル化への関与, MORITA KOTARO, HATANAKA YUKI, SHIMIZU NATSUMI, NISHIHARA TAKUSHI, TAKEMOTO ATSUSHI, HIGUCHI CHIKA, UCHIBORI SHO, AMANO TOMOKO, NAGAI KOHEI, KISHIGAMI SATOSHI, KATO HIROMI, MITANI TASUKU, HOSOI YOSHIHIKO, MATSUMOTO KAZUYA, 日本分子生物学会年会プログラム・要旨集(Web), 36th, 2P-0252 (WEB ONLY),   2013 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=201402296885395420
  • ベーチェット病患者末梢血単核球における発現蛋白質の網羅的検討, 吉岡拓也, 黒川真奈絵, 高桑由希子, 中野弘雅, 大岡正道, 飯塚進子, 佐藤利行, 永井宏平, 有戸光美, 岡本一起, 鈴木登, 加藤智啓, 日本リウマチ学会総会・学術集会・国際リウマチシンポジウムプログラム・抄録集, 56th-21st, 670, 670,   2012 03 19 , http://jglobal.jst.go.jp/public/201202278998589881
  • 翻訳後修飾プロテオミクスによる関節リウマチ特異的アセチル化蛋白質の解析, 有戸光美, 永井宏平, 高桑由希子, 大岡正道, 黒川真奈絵, 佐藤利行, 飯塚進子, 岡本一起, 加藤智啓, 日本リウマチ学会総会・学術集会・国際リウマチシンポジウムプログラム・抄録集, 56th-21st, 554, 554,   2012 03 19 , http://jglobal.jst.go.jp/public/201202284622768319
  • In vitroシトルリン化を用いたRAにおける新規シトルリン化抗原の同定, KUROKAWA MANAE, TAKENOUCHI KENJI, NAKAMURA YO, OKAMOTO KAZUKI, ARITO MITSUMI, SATO TOSHIYUKI, NAGAI KOHEI, YUDO KAZUO, KATO TOMOHIRO, 日本リウマチ学会総会・学術集会・国際リウマチシンポジウムプログラム・抄録集, 56th-21st, 343,   2012 03 19 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=201202209223124015
  • 関節リウマチにおける糖蛋白質の糖鎖構造の変異, SATO TOSHIYUKI, TAKAKUWA YUKIKO, OOKA SEIDO, NAGAI KOHEI, ARITO MITSUMI, IIZUKA MICHIKO, KUROKAWA MANAE, OKAMOTO KAZUKI, KATO TOMOHIRO, 日本リウマチ学会総会・学術集会・国際リウマチシンポジウムプログラム・抄録集, 56th-21st, 486,   2012 03 19 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=201202248357251841
  • NF‐κBを直接阻害する新しい抗炎症薬の作用機序, MITSUI HIROYUKI, OKAMOTO KAZUKI, KUROKAWA MANAE, ARITO MITSUMI, NAGAI KOHEI, SATO TOSHIYUKI, IIZUKA MICHIKO, YUDO KAZUO, BEPPU MOROE, KATO TOMOHIRO, 日本リウマチ学会総会・学術集会・国際リウマチシンポジウムプログラム・抄録集, 56th-21st, 325,   2012 03 19 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=201202235158948464
  • 自己抗原蛋白質U1‐small unclear ribonucleoprotein 68K subunitにおける疾患特異的翻訳後修飾の解析, NAGAI KOHEI, 聖マリアンナ医科大学雑誌, 39, 4, 284,   2012 03 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=201202266353662128
  • Comprehensive analysis of protein expression in peripheral blood mononuclear cells from patients with Behcet's disease, Kurokawa Manae, Suzuki, Suematsu Naoya, Okamoto Kazuki, Suzuki Noboru, Ozaki Syoichi, Kato Tomohiro, Yoshioka Takuya, Sato Toshiyuki, Iiduka Nobuko, Arito Mitsumi, Takakuwa Yukiko, Nakano Hiromasa, Ooka Seido, Nagai Kouhei, Abstracts for Annual Meeting of Japanese Proteomics Society, 2012, 0, 142, 142,   2012 , http://ci.nii.ac.jp/naid/130005454508
  • Comparative proteomic analysis of neutrophils from patients with microscopic polyangiitis and granulomatosis with polyangiitis., Uchida Teisuke, suematsu naoya, okamoto kazuki, ozaki shouichi, kato tomohiro, nagai kouhei, sato toshiyuki, iizuka nobuko, arito mitsumi, takakuwa yukiko, oooka seido, nakano hiromasa, kurokawa manae, Abstracts for Annual Meeting of Japanese Proteomics Society, 2012, 0, 147, 147,   2012 , http://ci.nii.ac.jp/naid/130005454504
  • Protein profiles of glomeruli obtained by micro-sieving in patients with IgA nephropathy, Kojima Shigeki, Yasuda Takashi, Kimura Kenjiro, Kato Tomohiro, Koitabashi Kenichiro, Iizuka Nobuko, Okamoto Kazuki, Arito Mitsumi, Sato Toshiyuki, Nagai Kouhei, Kurokawa(Suzuki) Manae, Suematsu Naoya, Abstracts for Annual Meeting of Japanese Proteomics Society, 2012, 0, 156, 156,   2012 , http://ci.nii.ac.jp/naid/130005454483
  • ANCA関連血管炎患者における好中球ミエロペルオキシダーゼの酸化修飾の変化, NAGAI KOHEI, UCHIDA TEISUKE, TAKAKUWA YUKIKO, OOKA SEIDO, ARITO MITSUMI, SATO TOSHIYUKI, KUROKAWA MANAE, OKAMOTO KAZUKI, SUEMATSU NAOYA, KATO TOMOHIRO, 日本生化学会大会(Web), 85th, 2P-250 (WEB ONLY),   2012 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=201402236548082906
  • 関節リウマチにおける糖蛋白質の糖鎖構造変異, SATO TOSHIYUKI, OOKA SEIDO, TAKAKUWA YUKIKO, ARITO MITSUMI, IIZUKA NOBUKO, NAGAI KOHEI, KUROKAWA MANAE, OKAMOTO KAZUKI, SUEMATSU NAOYA, KATO TOMOHIRO, 日本生化学会大会(Web), 85th, 2P-237 (WEB ONLY),   2012 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=201402230127648910
  • マウス初期胚におけるmajor ZGAでのユビキチン・プロテアソーム経路の関与, HIGUCHI CHIKA, SHIMIZU NATSUMI, HATANAKA YUKI, SHIN SEUNG-WOOK, NISHIKAWA SATOSHI, NISHIHARA TAKUJI, KATO RIE, TAKEMOTO ATSUSHI, MORITA KOTARO, NAGAI KOHEI, AMANO TOMOKO, KISHIGAMI SATOSHI, HOSOI YOSHIHIKO, MATSUMOTO KAZUYA, 日本分子生物学会年会プログラム・要旨集(Web), 35th, 3P-0489 (WEB ONLY),   2012 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=201302269789768190
  • ウシ僧帽筋のプロテオーム解析による筋内脂肪量に関わる新規タンパク質バイオマーカー候補の同定, TAKEMOTO ATSUSHI, NAGAI KOHEI, IKEGAMI HARUKA, HIGUCHI CHIKA, MORITA KOTARO, KOBAYASHI EIJI, MATSUMOTO KAZUYA, 日本分子生物学会年会プログラム・要旨集(Web), 35th, 4P-0030 (WEB ONLY),   2012 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=201302239550782089
  • マウス初期胚におけるユビキチン・プロテアソーム経路の転写制御への関与, SHIMIZU NATSUMI, HATANAKA YUKI, HIGUCHI CHIKA, SHIN SEUNG-WOOK, NISHIKAWA SATOSHI, NISHIHARA TAKUJI, KATO RIE, TAKEMOTO ATSUSHI, MORITA KOTARO, NAGAI KOHEI, AMANO TOMOKO, KISHIGAMI SATOSHI, ANZAI MASAYUKI, HOSOI YOSHIHIKO, MATSUMOTO KAZUYA, 日本分子生物学会年会プログラム・要旨集(Web), 35th, 3P-0488 (WEB ONLY),   2012 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=201302205140673647
  • 電気泳動を基礎にした臨床プロテオミクス最近の話題 消化器領域におけるプロテオミクスの応用, 初谷 守朗, 黒川 真奈絵, 永井 宏平, 有戸 光美, 末松 直也, 岡本 一起, 伊東 文生, 加藤 智啓, 生物物理化学, 55, Suppl., 9, 9,   2011 11
  • 自己抗原蛋白質U1‐small nuclear ribonucleoprotein 68k subunitにおける疾患特異的翻訳後修飾の解析, NAGAI KOHEI, 聖マリアンナ医科大学雑誌, 39, 2/3, 163,   2011 11 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=201102201744571184
  • 関節リウマチにおける糖タンパク質の糖鎖構造変異, 佐藤利行, 大岡正道, 高桑由希子, 永井宏平, 有戸光美, 飯塚進子, 黒川真奈絵, 岡本一起, 末松直也, 加藤智啓, 生化学, 84回, ROMBUNNO.3P-0111, 0111,   2011 09 , http://jglobal.jst.go.jp/public/201202219545052430
  • 自己免疫疾患患者の末梢血単核球における自己抗原U1‐small nuclear ribonucleoprotein 68k subunitの翻訳後修飾の変化, 永井宏平, 有戸光美, 高桑由希子, 大岡正道, 佐藤利行, 黒川真奈絵, 岡本一起, 内田貞輔, 末松直也, 加藤智啓, 生化学, 84回, ROMBUNNO.4P-0513, 0513,   2011 09 , http://jglobal.jst.go.jp/public/201202237988205025
  • ステロイド剤の抗炎症作用にはグルココルチコイドレセプター・コアクティベーター(MTI‐II)が必要である, 岡本一起, 三井寛之, 末松直也, 黒川真奈絵, 有戸光美, 永井宏平, 佐藤利行, 飯塚進子, 遊道和雄, 加藤智啓, 生化学, 84回, ROMBUNNO.4P-0290, 0290,   2011 09 , http://jglobal.jst.go.jp/public/201202289135772035
  • 「アセチル化」プロテオミクスによる関節リウマチ関連分子の探索, 有戸光美, 永井宏平, 高桑由希子, 大岡正道, 黒川真奈絵, 岡本一起, 加藤智啓, 日本リウマチ学会総会・学術集会・国際リウマチシンポジウムプログラム・抄録集, 55th-20th, 500, 500,   2011 06 20 , http://jglobal.jst.go.jp/public/201102262145288759
  • MPO‐ANCAの対応抗原の翻訳後修飾の検討, 内田貞輔, 永井宏平, 黒川真奈絵, 岡本一起, 高桑由希子, 大岡正道, 有戸光美, 加藤智啓, 日本リウマチ学会総会・学術集会・国際リウマチシンポジウムプログラム・抄録集, 55th-20th, 501, 501,   2011 06 20 , http://jglobal.jst.go.jp/public/201102286199467256
  • 抗RNP抗体の対応抗原snRNP68kの翻訳後修飾の解析, 永井宏平, 黒川真奈絵, 内田貞輔, 高桑由希子, 大岡正道, 有戸光美, 岡本一起, 加藤智啓, 日本リウマチ学会総会・学術集会・国際リウマチシンポジウムプログラム・抄録集, 55th-20th, 320,   2011 06 20 , http://jglobal.jst.go.jp/public/201102210498196670
  • 関節リウマチ関連因子Anx7の解析, 有戸光美, 松尾光祐, 野寄浩司, 中村洋, 黒川真奈絵, 増子佳世, 永井宏平, 岡本一起, 遊道和雄, 別府諸兄, 齋藤知行, 加藤智啓, 日本リウマチ学会総会・学術集会・国際リウマチシンポジウムプログラム・抄録集, 55th-20th, 331,   2011 06 20 , http://jglobal.jst.go.jp/public/201102261055728427
  • 核内受容体コアクティベーター(MTI‐II)はステロイドホルモンの抗炎症作用に関与する, 岡本一起, 佐藤利行, 永井宏平, 有戸光美, 黒川真奈絵, 末松直也, 遊道和雄, 礒橋文秀, 加藤智啓, ビタミン, 85, 4, 251, 251,   2011 04 25 , http://jglobal.jst.go.jp/public/201102217855250550
  • これから期待できるバイオマーカー 血清ペプチドミクス解析による新規腎疾患関連マーカーの探索, 永井宏平, 木村健二郎, 加藤智啓, 腎と透析, 70, 2, 224, 228,   2011 02 25 , http://jglobal.jst.go.jp/public/201102214763146820
  • 難治性血管炎に関する調査研究 MPO‐ANCA関連血管炎患者における自己抗原MPOの翻訳後修飾の検討, 加藤智啓, 内田貞輔, 永井宏平, 黒川真奈絵, 佐藤利行, 有戸光美, 岡本一起, 末松直也, 尾崎承一, 大岡正道, 高桑由希子, 遊道和雄, 難治性血管炎に関する調査研究 平成22年度 総括・分担研究報告書, 69, 73,   2011 , http://jglobal.jst.go.jp/public/201102299668120879
  • 消化器領域におけるプロテオミクスの応用, 初谷守朗, 黒川真奈絵, 永井宏平, 有戸光美, 末松直也, 岡本一起, 伊東文生, 加藤智啓, 生物物理化学(Web), 55, Suppl, S9(J-STAGE),   2011 , 10.2198/sbk.55.s9, http://jglobal.jst.go.jp/public/201202219521582358
  • Proteomic analysis of neutrophils of patient with ANCA-associated vasculitis using 2D-DIGE, Uchida Teisuke, Kato Tomohiro, Nagai Kouhei, Arito Mitsumi, Sato Toshiyuki, Takakuwa Yukiko, Ooka Seido, Suematsu Naoya, Kurokawa Manae, Okamoto Kazuki, Abstracts for Annual Meeting of Japanese Proteomics Society, 2011, 0, 153, 153,   2011 , http://ci.nii.ac.jp/naid/130005454351
  • 慢性C型肝炎患者におけるpeg化インターフェロンαおよびリバビリンの治療効果の、血清ペプチドミクスによる予測(Prediction of the effect of pegylated interferon alpha and ribavirin therapy by serum peptidomics in patients with chronic hepatitis C), 野口 陽平, 黒川 真奈絵, 奥瀬 千晃, 松本 伸行, 松永 光太郎, 永井 宏平, 有戸 光美, 佐藤 利行, 末松 直也, 岡本 一起, 伊東 文生, 加藤 智啓, 日本生化学会大会・日本分子生物学会年会合同大会講演要旨集, 83回・33回, 4P, 0198,   2010 12
  • 腎生検1検体から単離した糸球体抽出タンパク質の2次元電気泳動~糸球体腎炎研究への応用~, 小板橋賢一郎, 岡本一起, 有戸光美, 佐藤利行, 永井宏平, 黒川真奈絵, 末松直也, 安田隆, 木村健二郎, 加藤智啓, 生化学, 83回・33回, ROMBUNNO.4P-0195, 0195,   2010 12 , http://jglobal.jst.go.jp/public/201002265221723160
  • 関節リウマチ関連分子アネキシンVII(Anx7)の機能解析(Functional analysis of Rheumatoid Arthritis-related molecule, Annexin VII (Anx7)), 有戸 光美, 松尾 光祐, 黒川 真奈絵, 永井 宏平, 増子 佳世, 岡本 一起, 末松 直也, 加藤 智啓, 日本生化学会大会・日本分子生物学会年会合同大会講演要旨集, 83回・33回, 3P, 1056,   2010 12
  • 新規の核内レセプター・コアクティベーター(MTI‐II)タンパク質の細胞内への導入, 三井寛之, 岡本一起, 末松直也, 黒川真奈絵, 有戸光美, 永井宏平, 佐藤利行, 遊道和雄, 別府諸兄, 加藤智啓, 生化学, 83回・33回, ROMBUNNO.1P-0348, 0348,   2010 12 , http://jglobal.jst.go.jp/public/201002251689092620
  • グルココルチコイドレセプター・コアクティベーター(MTI‐II)による抗炎症作用, 岡本一起, 三井寛之, 末松直也, 黒川真奈絵, 有戸光美, 永井宏平, 佐藤利行, 遊道和雄, 加藤智啓, 生化学, 83回・33回, ROMBUNNO.1P-0347, 0347,   2010 12 , http://jglobal.jst.go.jp/public/201002273709911855
  • 関節リウマチ特異的に糖鎖構造変異を有する糖タンパク質の探索, 佐藤利行, 有戸光美, 永井宏平, 黒川真奈絵, 岡本一起, 末松直也, 加藤智啓, 生化学, 83回・33回, ROMBUNNO.3P-1060, 1060,   2010 12 , http://jglobal.jst.go.jp/public/201002276615059607
  • 自己抗原U1‐small nuclear ribonucleoprotein 68k subunitにおける疾患特異的翻訳後修飾亜型の解析, 永井宏平, 黒川真奈絵, 岡本一起, 内田貞輔, 高桑由希子, 大岡正道, 有戸光美, 佐藤利行, 末松直也, 加藤智啓, 生化学, 83回・33回, ROMBUNNO.3P-1062, 1062,   2010 12 , http://jglobal.jst.go.jp/public/201002282614534816
  • マウス初期胚における発生制御タンパク質の解析, 野老 美紀子, 川澄 みゆり, 永井 宏平, 池上 春香, 申 承旭, 西川 慧, 畑中 勇輝, 西原 卓志, 天野 朋子, 三谷 匡, 加藤 博巳, 安齋 政幸, 岸上 哲士, 佐伯 和弘, 細井 美彦, 入谷 明, 松本 和也, The Journal of Reproduction and Development, 56, Suppl., j120, j120,   2010 08 , 10.14882/jrds.103.0.124.0
  • Serum peptidomics, Kouhei Nagai, Nagayuki Kaneshiro, Kenjiro Kimura, Tomohiro Kato, Japanese Journal of Nephrology, 52, 475, 479,   2010 06 25 , http://www.scopus.com/inward/record.url?partnerID=HzOxMe3b&scp=77953774247&origin=inward
  • Serum peptidomics, NAGAI Kouhei, KANESHIRO Nagayuki, KIMURA Kenjiro, KATO Tomohiro, The Japanese journal of nephrology, 52, 4, 475, 479,   2010 05 25 , http://ci.nii.ac.jp/naid/10026409231
  • 血清ペプチドプロファイルによるペグインターフェロン・リバビリン併用療法のC型肝炎治療効果予測, 野口陽平, 黒川真奈絵, 奥瀬千晃, 松本伸行, 松永光太郎, 永井宏平, 有戸光美, 佐藤利行, 増子佳世, 末松直也, 岡本一起, 伊東文生, 加藤智啓, 日本消化器病学会雑誌, 107, 臨増総会, A271, A271,   2010 03 , http://jglobal.jst.go.jp/public/201002259903760857
  • プロテオミクスによるマウス着床前期胚の発生関連タンパク質の解析, 野老 美紀子, 川澄 みゆり, 永井 宏平, 池上 春香, 申 承旭, 西川 慧, 李 香欣, 畑中 勇輝, 天野 朋子, 三谷 匡, 加藤 博己, 安齋 政幸, 岸上 哲士, 佐伯 和弘, 細井 美彦, 入谷 明, 松本 和也, 日本畜産学会大会講演要旨集, 111回, 78, 78,   2009 09
  • 新規の核内レセプター・コアクティベーター(MTI‐II)の立体構造と転写促進活性, 岡本一起, 末松直也, 増子佳世, 黒川真奈絵, 有戸光美, 永井宏平, 遊道和雄, 加藤智啓, 生化学, 82回, 4T5p, 5,   2009 09 , http://jglobal.jst.go.jp/public/200902214009702859
  • 顕微鏡的多発血管炎患者血清ペプチドの網羅的探索, 高桑由希子, 黒川真奈絵, 大岡正道, 永井宏平, 有戸光美, 増子佳世, 末松直也, 岡本一起, 尾崎承一, 加藤智啓, 生化学, 82回, 3P, 713,   2009 09 , http://jglobal.jst.go.jp/public/200902252635520184
  • 末梢血単核球のプロテオーム解析による潰瘍性大腸炎とクローン病の鑑別診断, 初谷守朗, 黒川真奈絵, 紅露剛史, 永井宏平, 有戸光美, 増子佳世, 末松直也, 岡本一起, 伊東文生, 加藤智啓, 生化学, 82回, 2P, 348,   2009 09 , http://jglobal.jst.go.jp/public/200902257195829694
  • 関節リウマチ関連分子アネキシンA7の機能解析, 有戸光美, 松尾光祐, 黒川真奈絵, 永井宏平, 岡本一起, 増子佳世, 末松直也, 加藤智啓, 生化学, 82回, 3T15p, 7,   2009 09 , http://jglobal.jst.go.jp/public/200902298589959044
  • 多変量解析OPLS‐DAを用いたIgA腎症の血清ペプチドマーカーの探索, 永井宏平, 金城永幸, XIANG Yang, 黒川真奈絵, 岡本一起, 有戸光美, 増子佳世, 遊道和雄, 安田隆, 末松直也, 木村健二郎, 加藤智啓, 生化学, 82回, 4P, 685,   2009 09 , http://jglobal.jst.go.jp/public/200902214015772052
  • 片側内耳破壊後の前庭代償におけるラット小脳片葉タンパク質のプロテオーム解析, 深澤雅彦, 岡本一起, 中村学, 永井宏平, 有戸光美, 黒川真奈絵, 増子佳世, 末松直也, 肥塚泉, 加藤智啓, 生化学, 82回, 4P, 437,   2009 09 , http://jglobal.jst.go.jp/public/200902278905135309
  • プロテオミクスを用いたマウス初期胚における発生関連タンパク質の解析, 野老 美紀子, 川澄 みゆり, 永井 宏平, 池上 春香, 申 承旭, 西川 慧, 李 香欣, 畑中 勇輝, 天野 朋子, 三谷 匡, 加藤 博己, 安齋 政幸, 岸上 哲士, 佐伯 和弘, 細井 美彦, 入谷 明, 松本 和也, The Journal of Reproduction and Development, 55, Suppl., j142, j142,   2009 08 , 10.14882/jrds.102.0.1075.0
  • LHは卵巣内で一過性のタンパク質発現を誘起する 外因性ゴナドトロピン制御下におけるマウス卵巣のプロテオーム解析, 佐藤 学, 森本 義晴, 野老 美紀子, 申 承旭, 西川 慧, 畑中 勇輝, 佐伯 和弘, 細井 美彦, 入谷 明, 松本 和也, 池上 春香, 永井 宏平, 園 陽平, 福田 愛作, 日本生殖医学会雑誌, 54, 3, 119, 120,   2009 07
  • ビタミンB6酵素の転写を促進する核内受容体コアクティベーター(MTI‐II)のホモログタンパク質(prothymosin)の活性, 岡本一起, 永井宏平, 有戸光美, 黒川真奈絵, 増子佳世, 末松直也, 遊道和雄, 礒橋文秀, 加藤智啓, ビタミン, 83, 4, 177,   2009 04 25 , http://jglobal.jst.go.jp/public/200902250448911355
  • Proteome analysis of mouse preimplantation embryos, TOKORO Mikiko, KAWASUMI Miyuri, NAGAI Kohei, IKEGAMI Haruka, SATOH Manabu, SHIN Seung-Wook, NISHIKAWA Satoshi, HATANAKA Yuki, AMANO Tomoko, MITANI Tasuku, KATO Hiromi, ANZAI Masayuki, KISHIGAMI Satoshi, SAEKI Kazuhiro, HOSOI Yoshihiko, IRITANI Akira, MATSUMOTO Kazuya, Journal of mammalian ova research = 日本哺乳動物卵子学会誌, 26, 2,   2009 04 01 , http://ci.nii.ac.jp/naid/10024965056
  • 14日齢ウシ体細胞核移植胚のプロテオーム解析, MATSUI TAKANORI, IDETA ATSUSHI, NAGAI KOHEI, IKEGAMI HARUKA, URAKAWA MAMI, IWAMOTO TAISAKU, TANIGUCHI TOSHIHITO, HOSOI YOSHIHIKO, MATSUMOTO KAZUYA, IRIYA AKIRA, AOYAGI TAKAHITO, SAEKI KAZUHIRO, 日本畜産学会大会講演要旨, 110th, 8, 8,   2009 03 27 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=200902244608882250
  • マウス着床前期胚における大規模プロテオーム解析, 野老 美紀子, 川澄 みゆり, 永井 宏平, 池上 春香, 佐藤 学, 申 承旭, 西川 慧, 清水 なつみ, 天野 朋子, 三谷 匡, 加藤 博己, 安齋 政幸, 岸上 哲士, 佐伯 和弘, 細井 美彦, 入谷 明, 松本 和也, The Journal of Reproduction and Development, 54, Suppl., j95, j95,   2008 08 , 10.14882/jrds.101.0.507.0
  • 14日齢ウシ体細胞核移植胚におけるプロテオーム解析, MATSUI TAKANORI, IDETA ATSUSHI, NAGAI KOHEI, IKEGAMI HARUKA, URAKAWA MAMI, IWAMOTO TAISAKU, TANIGUCHI TOSHIHITO, HOSOI YOSHIHIKO, MATSUMOTO KAZUYA, IRIYA AKIRA, AOYAGI TAKAHITO, SAEKI KAZUHIRO, 日本畜産学会大会講演要旨, 109th, 11,   2008 03 27 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=200902262436230482
  • A System for Brand Beef-cattle (Hidagyu) Breeders to Support Selecting Well-Suited Sires, SHI Linjing, INOUE Etsuko, YOSHIHIRO Takuya, NAGAI Kouhei, IKEGAMI Haruka, KOBAYASHI Naohiko, MATSUMOTO Kazuya, NAKAGAWA Masaru, 全国大会講演論文集, 70, 0, 677, 678,   2008 03 13 , http://ci.nii.ac.jp/naid/110006864668
  • マウス着床前期胚における大規模プロテオーム解析, TOKORO MIKIKO, KAWASUMI MIYURI, NAGAI KOHEI, IKEGAMI HARUKA, SATO MANABU, SHIN SEUNG-WOOK, NISHIKAWA SATOSHI, AMANO TOMOKO, MITANI TASUKU, KATO HIROMI, ANZAI MASAYUKI, KISHIGAMI SATOSHI, SAEKI KAZUHIRO, HOSOI YOSHIHIKO, IRITANI AKIRA, MATSUMOTO KAZUYA, 生化学, 4P-0894,   2008 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=200902223409224866
  • 過排卵処理したマウス卵巣におけるプロテオーム解析, 松本 和也, 池上 春香, 永井 宏平, 園 陽平, 野老 美紀子, 申 承旭, 松岡 俊樹, 三谷 匡, 加藤 博己, 岸上 哲士, 佐伯 和弘, 細井 美彦, 入谷 明, 日本胚移植学雑誌, 30, 1, 55, 55,   2008 01
  • 飛騨牛由来白色脂肪組織の大規模プロテオーム解析, 池上春香, 永井宏平, 吉廣卓哉, 園陽平, 小林直彦, 松橋珠子, 大谷健, 中川優, 森本康一, 松本和也, 日本畜産学会大会講演要旨, 108th, 4,   2007 09 26 , http://jglobal.jst.go.jp/public/200902285597113656
  • マウス排卵卵子における網羅的タンパク質発現(プロテオーム)解析, 野老 美紀子, 川澄 みゆり, 永井 宏平, 池上 春香, 申 承旭, 松岡 俊樹, 佐藤 学, 天野 朋子, 三谷 匡, 加藤 博巳, 安齋 政幸, 岸上 哲士, 佐伯 和弘, 細井 美彦, 入谷 明, 松本 和也, The Journal of Reproduction and Development, 53, Suppl., j137, j137,   2007 09 , 10.14882/jrds.100.0.12047.0
  • マウスMII期卵母細胞における網羅的タンパク質発現(プロテオーム)解析, TOKORO MIKIKO, KAWASUMI MIYURI, NAGAI KOHEI, IKEGAMI HARUKA, SHIN SEUNG-WOOK, MATSUOKA TOSHIKI, SATO MANABU, AMANO TOMOKO, MITANI TASUKU, KATO HIROMI, ANZAI MASAYUKI, KISHIGAMI SATOSHI, SAEKI KAZUHIRO, HOSOI YOSHIHIKO, IRITANI AKIRA, MATSUMOTO KAZUYA, 生化学, 2P-1307,   2007 , http://jglobal.jst.go.jp/detail.php?from=API&JGLOBAL_ID=200902268338533254
  • マウス卵巣におけるプロテオーム解析の確立, 上中 崇裕, 永井 宏平, 池上 春香, 松本 和也, 天野 朋子, 佐伯 和弘, 細井 美彦, 入谷 明, The Journal of Reproduction and Development, 51, Suppl., j82, j82,   2005 08 , 10.14882/jrds.98.0.60.0
  • COMPARATIVE PROTEOMIC ANALYSIS OF NEUTROPHILS FROM PATIENTS WITH MICROSCOPIC POLYANGIITIS AND GRANULOMATOSIS WITH POLYANGIITIS, T. Uchida, K. Nagai, T. Sato, N. Iizuka, M. Arito, Y. Takakuwa, H. Nakano, S. Ooka, M. Kurokawa, N. Suematu, K. Okamoto, S. Ozaki, T. Kato, ANNALS OF THE RHEUMATIC DISEASES, 72, 832, 832,   2013 06
  • Comprehensive Analysis of Protein Expression in Peripheral Blood Mononuclear Cells From Patients with Behcet's Disease., Takuya Yoshioka, Manae Kurokawa, Yukiko Takakuwa, Hiromasa Nakano, Seido Ooka, Nobuko Iizuka, Toshiyuki Sato, Mitsumi Arito, Kouhei Nagai, Kazuki Okamoto, Naoya Suematsu, Noboru Suzuki, Shoichi Ozaki, Tomohiro Kato, ARTHRITIS AND RHEUMATISM, 64, 10, S78, S79,   2012 10

Research Grants & Projects

  • Ministry of Education, Culture, Sports, Science and Technology, Grants-in-Aid for Scientific Research(若手研究(B)), Establishment of maker for early diagnosis of systemic autoimmune diseases based on de-phosphorylation of autoimmogen U1-68k, In this study, we tried to establish an useful method for detecting a de-phosphorilated form of U1-68k autoimmunogen, which was found to be increased in peripheral blood mononuclear cells of patients of systemic lupus erythematosus or Mixed connective tissue diseases. As a substitution of a conventional 2D-western blot method, we examined three methods; (1) an immunological method, (2) a phos-tag electrophorsis, and (3)an isoelectric focusing electrophoresis-western blot. As a result, we found that the isoelectric focusing electrophoresis-western blot method is most suitable for the detection of the de-phosphorilated form of U1-68k. Moreover, we found that whole cell extract prepared by cell lysis buffer containing SDS can be used for detecting U1-68k. In conclusion, combination of isoelectric focusing electrophoresis-western blot and whole cell extract by SDS-containing cell lysis buffer can shorten analsys time from 5 days to 2.5 days.
  • Ministry of Education, Culture, Sports, Science and Technology, Grants-in-Aid for Scientific Research(若手研究(B)), The establishment of 'novel clinical examination for rheumatoid arthritis' by focusing on acetylation of proteins, Post-translational modifications (PTMs) are often critical for diagnosis of diseases. Thereby, we here tried to elucidate alteration of PTMs in RA, focusing on acetylation in this study.We applied acetyl-proteomics to peripheral blood mononuclear cells (PBMCs) to elucidate PTM difference between patients with RA and healthy. Multiple proteins in PBMCs were highly acetylated in the RA groups. One of the proteins predominantly acetylated in the RA group was identified to be ENO1. Next, we tried to identified acetylated lysine residues, but we could not detect RA-specific acetylated peptides of ENO1. This would be technical limitation of mass spectrometric analysis at present. In the future, it would be needed to establish more effective methods to identify acetylated lysine residues. Next, we examined whether acetylation affect the antigenecity of ENO1 in patients with RA. As a result, the reactivity was not different between the acetylated ENO1 and the non-acetylated ENO1.
  • Andlysis of Protease-catalyzed coagulation of Soy protein