KINDAI UNIVERSITY


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MORIMOTO Koichi

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FacultyDepartment of Genetic Engineering / Graduate School of Biology-Oriented Science and Technology
PositionProfessor
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Commentator Guidehttps://www.kindai.ac.jp/meikan/339-morimoto-kouichi.html
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Last Updated :2020/09/02

Education and Career

Education

  •  - 1987 , Hokkaido University
  •  - 1987 , Hokkaido University, Graduate School, Division of Natural Science

Academic & Professional Experience

  •   2012 , Faculty of Biology-Oriented Science and Technology, Kindai University

Research Activities

Research Areas

  • Life sciences, Biomaterials
  • Life sciences, Biomedical engineering
  • Life sciences, Functional biochemistry
  • Life sciences, Applied biochemistry

Research Interests

  • Saccharina japonica, Molecular Biotechnology, Enzyme Chemistry, Immunotechnology

Published Papers

  • Latent nature of collagen in promoting three-dimensional adherent spheroid formation of fibroblasts, Koichi Morimoto, Saori Kunii, Materialia, Materialia, 8, Dec. 01 2019
    Summary:© 2019 Fibroblast migration in connective tissues is important for remodeling and renewal. Fibroblast spheroids are useful for understanding biological activity in physiologically relevant environments. Recently, we developed low adhesive scaffold collagen (LASCol) from porcine type I collagen by actinidain hydrolysis and found that fibroblasts cultured on the LASCol matrix exhibited spontaneous, rapid spheroid formation. In this study, the formation and properties of fibroblast spheroids were characterized in detail. The surfaces of fibroblast spheroids were clearly distinct from those of monolayer fibroblasts, including differences in the number and shape of membrane protrusions. Interestingly, the moving speed of spheroid on the LASCol matrix was maintained at approximately 0.9 µm/min, even after 24 h of incubation. Our results also indicated that cell proliferation became arrested in spheroids while the relative expression levels of the genes encoding fibronectin, integrin β3, bFGF, PDGF-A, and TGF-β1 increased. LASCol is useful for obtaining adherent spheroids for tissue engineering and drug screening. We demonstrated that collagen has a latent nature in forming three-dimensional adherent spheroids of fibroblasts and in promoting the essential functions of fibroblasts.
  • 食品・バイオにおける最新の酵素応用 I コラーゲン酵素分解物の再生医療および食品科学への応用, 森本康一, 國井沙織, Bio Industry, Bio Industry, 36(5), 74‐81, May 2019
  • Structural analyses of type I collagen fibrils for application to tissue engineering, Koichi Morimoto, Saori Kunii, Type I Collagen: Molecular Structure, Applications in Tissue Engineering and Role in Human Disorders, Type I Collagen: Molecular Structure, Applications in Tissue Engineering and Role in Human Disorders, 73 - 85, Jul. 01 2015
    Summary:© 2015 by Nova Science Publishers, Inc. Type I collagen molecules spontaneously form insoluble fibrils at physiological buffers and at neutral pH. Insoluble fibrils play an important role in not only structure stabilization in tissue but also maintenance of each cell. Recently, collagen has been used as one of cell scaffolds in regenerative medicine. In cell culture conditions, the structure of molecule in collagen fibrils would be different from that of collagen monomer. Then, the study in ultrastructure of fibril is more important than that of monomer to understand cell behaviors. However, the structural information is not well understood yet. In this Chapter, we report on the helicity of collagen molecule in fibrils by circular dichroism spectroscopy and the thermograph of fibrils by differential scanning calorimetry. Furthermore, we observed the appearance of mouse neutrophil on collagen scaffolds showing imperfect fibrils. Our data indicated that the ultrastructure of collagen molecule depends on monomer or fibrils. We suggest that the formation of collagen fibrils prior to cell seeding is necessary for the interaction between cells and extracellular matrices.
  • Examination of effective culture methods for rabbit preantral follicles, Hironobu Sugimoto, Yuki Miyamoto, Yoko Tsuji, Kouichi Morimoto, Takeshi Taniguchi, Yoshiharu Morimoto, Yoshihiko Hosoi, Journal of Mammalian Ova Research, Journal of Mammalian Ova Research, 26(4), 221 - 226, Oct. 2009
    Summary:To determine an effective in vitro culture method of rabbit immature ovarian follicles, preantral follicles (200 to 299 μm in diameter) were cultured by the whole follicle culture method and by the open type culture method. The use of tuna matrix collagen (MC) as a medium material to maintain 3-dimensional structure of the follicles was also investigated for the open type culture method. In the whole follicle culture method, the diameter of the preantral follicles increased from 252.8 ± 2.7 μm to 395.6 ± 9.6 μm. However, culture for oocyte maturation was not possible because most of the oocytes had degenerated. In the open culture method, in which oocyte-granulosa cell complexes (OGs) collected from preantral follicles were cultured, the rates of the oocytes that resumed meiosis were 1.6%, 5.4%, and 73.7% in 0, 0.3, and 3 mg/ml of MC supplementation, respectively. Morphologically, the addition of MC was found to show a beneficial effect for maintenance of the three-dimensional structure of the follicles. In conclusion, open type culture with MC supplementation seems to be a most appropriate culture method for rabbit immature follicles.
  • 順相分配クロマトグラフィーによるニワトリI型コラーゲンのα1鎖とα2鎖の分離精製, 國井沙織, 森本康一, Memoirs of the School of Biology-Oriented Science and Technology of Kinki University, Memoirs of the School of Biology-Oriented Science and Technology of Kinki University, (22), 25 - 32, Sep. 30 2008
  • Microbiological evaluation of fruits certified as Specially Grown Agricutural products by an accerdited certification agency, 20, 1 - 8, Sep. 2007
  • Microbiological Evaluation of Fruits Certified as Specially Grown Agricultural Products by an Accredited Certification Agency, IZUMI H, Memoirs of the School of Biology-Oriented Science and Technology of Kinki University, Memoirs of the School of Biology-Oriented Science and Technology of Kinki University, (20), 1 - 8, Sep. 2007
  • Matrix metalloproteinase‐1(MMP‐1)分解によるI型コラーゲン会合体の構造解析, 森本康一, 國井沙織, Memoirs of the School of Biology-Oriented Science and Technology of Kinki University, Memoirs of the School of Biology-Oriented Science and Technology of Kinki University, 19(19), 57 - 65, Mar. 31 2007
    Summary:The degradation of type I collagen fibril in vivo is most important event in cell invasion, tissue repair, and organ morphogenesis. Matrix metalloproteinase-1 (MMP-1) is one of enzyme, which hydrolyzes triple-helical structure of the native collagen. The collagen fragments cleaved by MMP-1 are readily digested to small polypeptides. The enzyme specificity of bacterial MMP-1 would be different among the species. It also depends on molecular structure of the collagen self-assemblies. The digested fragments were investigated using SDS-polyacrylamide gel electrophoresis and reverse phase HPLC. Here we report that the structural difference between pepsin-treated collagen and actinidain-treated collagen had influence on the enzymatic activity of MMP-1. The study also shows that the degradation rate of tuna collagen was much faster than that of chicken collagen and MMP-ls produced from Clostridium histolyticum and Streptomyces were a minimal difference in specificity.
  • Comparison of biochemical properties of novel actinidain-hydrolyzed collagen and pepsin-hydrolyzed collagen, (10), 19 - 28, Mar. 2005
  • Preparation and structural analysis of actinidain-processed atelocollagen of yellowfin tuna (Thunnus albacares), Morimoto K, Kunii S, Hamano K, Tonomura B, Biosci. Biotechnol. Biochem., Biosci. Biotechnol. Biochem., 68(4), 861 - 867, Apr. 2004 , Refereed
  • The steady-state and the pre-steady-state kinetic analysis of the hydrolysis of N-α-Cbz-L-Lysine p-nitrophenyl ester catalysed by actinidain isozymes, Morimoto Koichi, Tamura Masaya, Tonomura Ben'ichiro, Memoirs of the School of Biology-Oriented Science and Technology of Kinki University, Memoirs of the School of Biology-Oriented Science and Technology of Kinki University, 10(10), 29 - 37, Jul. 30 2002
    Summary:The steady-state and the pre-steady-state kinetics of two actinidain isozymes, AC-1 and AC-2, were analyzed by stopped-flow method with N-alpha-carbo-benzoxy-L-lysine p-nitrophenyl ester (Z-Lys-ONp) as substrate. These isozymes were purified from kiwifruit (Actinidia deliciosa) by anion-exchange chromatography. The steady-state kinetic parameters, K_m and V, were determined. Rapid bursts observed in the pre-steady-state for two isozymes are consistent with a three-step mechanism involving an acyl-enzyme intermediate. The values of k_<cat>/K_m and k_<+2>/K_m^<acyl> ratios for AC-1 were estimated 1.83 and 8.63 μM^<-1>・s^<-1> respectively, and were two times greater than those for AC-2 (k_<cat>/K_m = 1.02, k_<+2>/K_m^<acyl> = 4.50 μM^<-1>・s^<-1>). For AC-1 and AC-2, the deacylation step is rate-limiting in catalysis, so k_<cat> is similar to k_<+3> under the experimental conditions.
  • The steady-state and pre-steady-state kinetic analysis of the hydrolysis of N-a-Cbz-L-Lysine p-nitrophenyl ester catalysed by actinidain isozymes, Mem. School, B.O.S.T. Kinki University, Mem. School, B.O.S.T. Kinki University, (10), 29 - 37, 2002
  • Method for the preparation of bispecific F(ab ')2 mu fragments from mouse monoclonal antibodies of the immunoglobulin M class and characterization of the fragments, K Morimoto, K Inouye, JOURNAL OF IMMUNOLOGICAL METHODS, JOURNAL OF IMMUNOLOGICAL METHODS, 224(1-2), 43 - 50, Apr. 1999
    Summary:Bispecific F(ab')2 mu, fragments (Bs F(ab')2 mu) binding simultaneously both sialyl Lewis A antigen (SLA) and human carcinoembryonic antigen (CEA) were prepared by disulfide bond exchange between F(ab')2 mu, fragments derived from IgM monoclonal antibodies (mAbs) against SLA and CEA, and were purified to homogeneity in a one-step procedure of hydrophobic interaction HPLC. The final yield of Bs F(ab')2 mu from F(ab')2 mu fragments was 70-78%, and the purity was higher than 98%. The immunoreactivities of the Bs F(ab')2 mu fragments against SLA and CEA were almost the same as those of the respective parental F(ab')2 mu, fragments. The dissociation constant (0.17 mu M) of the Bs F(ab')2 mu, for CEA was in good agreement with that of the parental F(ab')2 mu, fragments. Although the number of applications of IgM mAbs is restricted because of the large molecular mass and low solubility, Bs F(ab')2 mu might, nevertheless, be a useful tool for immunotherapy and immunodiagnosis. (C) 1999 Elsevier Science B.V. All rights reserved.
  • Flow cytometric analysis of sialyl Lewis A antigen on human cancer cells by using F(ab')(2 mu) fragments prepared from a mouse IgM monoclonal antibody, K Morimoto, K Inouye, CYTOTECHNOLOGY, CYTOTECHNOLOGY, 24(3), 219 - 226, 1997
    Summary:F(ab')(2) fragments, herein designated as F(ab')(2 mu) fragments, were prepared from a mouse IgM monoclonal antibody specific to sialyl Lewis A antigen. The fragments were applied to flow cytometry to analyze the antigen on human cancer cells. The binding of the fragments to the antigen-positive cells was stronger than that of the original IgM. The non-specific binding of the IgM antibody to the antigen-negative cells was much decreased by using the F(ab')(2 mu) fragments. These results indicate that the F(ab')(2 mu) fragments are more suitable than the original IgM monoclonal antibody in flow cytometric analysis.
  • Preparation of F(ab′)2μ fragments of IgM monoclonal antibodies, K. Morimoto, K. Inouye, Seikagaku, Seikagaku, 68, 42 - 45, Dec. 01 1996
  • Single-step purification of F(ab′)2μ fragments of mouse monoclonal antibodies (immunoglobulins M) by hydrophobic interaction high-performance liquid chromatography using TSKgel Ether-5PW, Kuniyo Inouye, Koichi Morimoto, Journal of Biochemical and Biophysical Methods, Journal of Biochemical and Biophysical Methods, 26(1), 27 - 39, 1993
    Summary:A procedure is described for preparation and single-step purification of F(ab′)2 fragments, herein designated as F(ab′)2μ′ from mouse monoclonal antibodies of the IgM class. Hydrophobic interaction high-performance liquid chromatography (HPLC) using TSKgel Ether-5PW was well applicable to the purification. The IgM was digested with pepsin at the pepsin-to-IgM ratio of 1:200 (w/w) in 100 mM citrate buffer (pH 4.2) at 37°C for 2 h. The digests were applied to the gel equilibrated with the buffer containing 1 M ammonium sulfate. F(ab′)2μ fragments were adsorbed onto the gel with the same buffer, and eluted by reducing the ammonium sulfate concentration to 0 M. The fraction containing F(ab′)2μ fragments was homogeneous (purity higher than 97%) by both sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel-filtration HPLC. The recovery of the antigen-binding site was 55-72%. The cycle time of the Ether-5PW HPLC was 40 min, and up to 98 mg F(ab′)2μ was purified in a cycle. This method could be suitable especially for large-scale purification of F(ab′)2μ fragments. The molecular mass of F(ab′)2μ was estimated to be 144-146 kDa. In comparison with IgM, F(ab′)2μ lost entirely the complement C1q binding activity, and the sugar content was greatly reduced. The binding of IgM with non-specific proteins turned to be negligible, when IgM was converted to F(ab′)2μ, suggesting that the fragments are useful for immunological application. © 1993.
  • Single-step purification of F(ab′)2 fragments of mouse monoclonal antibodies (immunoglobulins G1) by hydrophobic interaction high performance liquid chromatography using TSKgel Phenyl-5PW, Koichi Morimoto, Kuniyo Inouye, Journal of Biochemical and Biophysical Methods, Journal of Biochemical and Biophysical Methods, 24(1-2), 107 - 117, 1992
    Summary:Hydrophobic interaction high performance liquid chromatography (HPLC) using TSKgel Phenyl-5PW was applicable to single-step purification of F(ab′)2 fragments from pepsin digests of mouse monoclonal antibodies of IgG1 class. The digests were applied to the gel equilibrated with phosphate-buffered saline containing 1 M ammonium sulfate. F(ab′)2 fragments were adsorbed onto the gel using the same buffer, and eluted by reducing the ammonium sulfate concentration to 0 M. The fraction containing F(ab′)2 fragments was homogeneous (purity: higher than 98%) by both sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel filtration HPLC. The recovery of the antigen binding site was 42-58%. The cycle time of the Phenyl-5PW HPLC was 45 min, and F(ab′)2 of up to 2200 mg was purified in a cycle. This method could be useful especially for large scale purification of F(ab′)2 fragments. © 1992.
  • Recovery of active recombinant EGFP from the excrement of transgenic mice: A possible source of recombinant protein, Masateru Magotani, Aya Nakamura, Haruka Ikegami, Saori Kunii, Yuji Mishina, Koichi Morimoto, Satoshi Kishigami, Biochemical and Biophysical Research Communications, Biochemical and Biophysical Research Communications, 500(3), 817 - 823, Jun. 07 2018 , Refereed
    Summary:Transgenic animals provide attractive systems for the production of valuable recombinant proteins. Previous studies indicate that milk is a suitable source of secreted recombinant proteins. In the current study, we examine whether excrement can be another source of recombinant proteins by using transgenic mice ubiquitously-expressing green fluorescent protein (GFP) as a model. We found that the surface of excrement from GFP-transgenic mice was fluorescent, and the supernatant after centrifugation of an excrement suspension was rich in undegraded, actively fluorescing GFP. GFP was successfully purified from stool as a fluorescent 27 kDa protein by using a common procedure. Finally, we observed that the fluorescence of digested materials began in the ileum and persisted throughout the remainder of the digestive tract. Our results demonstrate that excrement, which is produced daily regardless of the sex or age of the animal, may be another feasible source of recombinant proteins. The preparation method is simple, economical, and noninvasive.
  • 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.
  • 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.
  • Characterization of Type I Collagen Fibril Formation Using Thioflavin T Fluorescent Dye, Koichi Morimoto, Kazuya Kawabata, Saori Kunii, Kaori Hamano, Takuya Saito, Ben'ichiro Tonomura, JOURNAL OF BIOCHEMISTRY, JOURNAL OF BIOCHEMISTRY, 145(5), 677 - 684, May 2009 , Refereed
    Summary:Collagen is composed of fibrils that are formed by self-assembly of smaller units, monomers which are triple-helical polypeptide. However, the mechanism of fibril formation at the level of individual molecules has remained to be clarified. We found that the fluorescence of thioflavin T, which has been widely used as a specific dye for amyloid fibrils, also increased by binding with fibrils of atelocollagen prepared from yellowfin tuna skin. There was a linear correlation between the fluorescence increase and the amount of atelocollagen within a collagen concentration range of 00.15 mg/ml at pH 6.5 with 50 M thioflavin T. In contrast, neither actinidain-processed collagen that keeps monomeric nature nor heat-denatured collagen could cause the fluorescence increase of thioflavin T at all. The relationship between the fluorescence increase and thioflavin T concentration was fit to a theoretical binary binding curve. An apparent dissociation constant, K-d, and a maximal fluorescence increase, F-max, were calculated at various pHs. The values of K-d and F-max were dependent on pH (K-d was 9.4 M at pH 6.5). The present finding demonstrates that thioflavin T specifically binds to collagen fibrils and may be used as a sensitive tool for the study of collagen structure.
  • 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.
  • Effects of high concentration of salts on the esterase activity and structure of a kiwifruit peptidase, actinidain, Koichi Morimoto, Erino Furuta, Hiroshi Hashimoto, Kuniyo Inouye, JOURNAL OF BIOCHEMISTRY, JOURNAL OF BIOCHEMISTRY, 139(6), 1065 - 1071, Jun. 2006 , Refereed
    Summary:Effects of salts on the activity and stability of actinidain were examined. With increasing salt concentration up to 0.5 M, the activity (k(cat)/K-m) for N-alpha-Cbz-L-lysine p-nitrophenyl ester decreased to 40% of that in the absence of salt. The inhibitor constant K-i of LiCl, NaCl, and KCl was 0.16-0.43 M. With 3 M KCl and NaCl, the specificity constant k(cat)/K-m recovered to 110 and 75%, respectively. No re-activation was observed with LiCl. The inhibition and re-activation were dependent on the changes in both K-m and k(cat), whereas no CD change was observed. The tryptophan fluorescence of actinidain was not affected by 0-0.5 M salt, but a considerable decrease in its intensity was observed with increasing salt concentration from 0.5 to 3.0 M. These results suggest that the inhibition observed with the lower salt concentration (< 0.5 M) is due to attenuation of the electrostatic interaction between the enzyme and substrate, and the higher concentration (0.5-3.0 M) induces structural change in the states of tryptophan residues, which is associated with the re-activation. Actinidain keeps considerably high activity and stability even in the presence of 3 M salts.
  • 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.
  • Protein adsorption on patterned hydroxyapatite thin films fabricated by pulsed laser deposition, M Kusunoki, M Kawasima, H Nishikawa, K Morimoto, T Hayami, S Hontsu, T Kawai, JAPANESE JOURNAL OF APPLIED PHYSICS PART 2-LETTERS & EXPRESS LETTERS, JAPANESE JOURNAL OF APPLIED PHYSICS PART 2-LETTERS & EXPRESS LETTERS, 44(8-11), L326 - L327, 2005
    Summary:Protein adsorption on hydroxyapatite (HAP) thin film was investigated before and after patterning. Hydroxyapatite thin film 100 nm thick was deposited by pulsed laser deposition. The film was patterned by photolithography and wet etching with HCl solution. Proteins (phospholyrase b, bovine serum albumin, and others) labeled with fluorescein isothiocyanate (FITC) were used as the reagent. After the HAP film was soaked in the reagent and washed with pure water, a conspicuous contrast in FITC was observed between the HAP pattern and the glass substrate (or photoresist). This behavior showed that the biocompatibility of the HAP thin film was not influenced by the patterning process. Our technique for HAP thin film is adaptable for applications involving biosensors as electronic devices and scaffolds for tissue culture.
  • Presence of protein complex is prerequisite for aragonite crystallization in the nacreous layer, A Matsushiro, T Miyashita, H Miyamoto, K Morimoto, B Tonomura, A Tanaka, K Sato, MARINE BIOTECHNOLOGY, MARINE BIOTECHNOLOGY, 5(1), 37 - 44, Jan. 2003
    Summary:We have isolated a protein complex from the nacreous layer of pearl beads and oyster shells. This complex was mainly composed of pearlin and pearl keratin. Addition of a minute amount of the complex to a calcium-carbonate-saturated solution containing Mg2+ induced aragonite crystallization. The complex was dissociated to individual components in the presence of EDTA and urea. Conversely, the complex was reconstituted from a mixture of components upon incubation with Ca2+ and urea. The mixture of the components was unable to induce aragonite crystallization, but the reconstituted complex recovered this capacity. Thus it is concluded that the complex is the indivisible functional unit required for aragonite crystallization.
  • A sensitive enzyme immunoassay of human thyroid-stimulating hormone (TSH) using bispecific F(ab')(2) fragments recognizing polymerized alkaline phosphatase and TSH, K Morimoto, K Inouye, JOURNAL OF IMMUNOLOGICAL METHODS, JOURNAL OF IMMUNOLOGICAL METHODS, 205(1), 81 - 90, Jun. 1997
    Summary:Bispecific F(ab')(2) fragments recognizing both human thyroid-stimulating hormone (TSH) and alkaline phosphatase (ALP) were prepared by disulfide bond exchange between F(ab')(2) fragments of IgGI monoclonal antibodies (mAbs) against TSH and ALP, and were purified to homogeneity by hydrophobic interaction HPLC, ALP was polymerized by glutaralde hyde, and a new sandwich enzyme-linked immunosorbent assay (ELISA) for TSH was developed by using the ALP polymers and bispecific F(ab')(2) fragments against TSH and ALP. In this assay, the preparation of covalently linked enzyme-mAb conjugates was not needed, and the interaction of mAb with non-specific proteins was greatly reduced by the use of F(ab')(2) fragments. The sensitivity for TSH was shown to increase in proportion to the degree of polymerization of ALP, and the lower detection limit obtained with the ALP trimer was 0.5 mu U/ml. The sensitivity was 30 times or more higher than that of the conventional ELISA using covalently linked enzyme-mAb conjugates. The use of bispecific F(ab')(2) permits the use of monomers and polymers of the signal enzyme and, thereby, regulates the sensitivity of the assay system. (C) 1997 Elsevier Science B.V.
  • Preparation of F(ab′)2μ fragments from rat IgM monoclonal antibodies and their application to the enzyme immunoassay of mouse interleukin-6, Kuniyo Inouye, Koichi Morimoto, Journal of Immunological Methods, Journal of Immunological Methods, 171(2), 239 - 244, May 16 1994
    Summary:F(ab′)2 fragments, herein designated as F(ab′)2μ, were prepared from rat IgM monoclonal antibodies (mAbs). The IgM was digested at a pepsin-to-IgM ratio of 1:200 (w/w) in 100 mM citrate buffer (pH 4.5) at 37°C for 2 h. During digestion, the light (L) chain (27 kDa) of IgM remained undegraded, whereas the heavy (H) chain disappeared and two new bands of 44 and 48 kDa appeared. The digests were fractionated by means of hydrophobic interaction HPLC with TSKgel Phenyl-5PW. The fraction containing F(ab′)2μ was homogeneous and the recovery of antigen-binding activity was 41-52%. The molecular mass of F(ab′)2μ was estimated to be 147-153 kDa, and we concluded that the fragment was composed of two truncated H chains and two intact L chains. F(ab′)2μ was used in an enzyme immunoassay of mouse interleukin-6 and the interaction of IgM with non-specific proteins was greatly reduced, when it was converted to F(ab′)2μ fragments. © 1994.

Conference Activities & Talks

  • 細胞低接着性コラーゲンが未分化維持培養におけるヒトiPS細胞の形態および未分化性に及ぼす影響, 米澤優希, 大貫義嗣, 國井沙織, 伊田寛之, 平岡陽介, 森本康一, 黒澤尋, 日本再生医療学会総会(Web),   2019
  • 細胞低接着性コラーゲンを足場とした神経細胞の突起伸長, 中野法彦, 兼清健志, 國井沙織, 森本康一, 尾前薫, 井出千束, 日本再生医療学会総会(Web),   2019
  • 骨組織から抽出した細胞低接着性コラーゲンの骨芽細胞分化誘導, 國井沙織, 堀内喜高, 伊田寛之, 平岡陽介, 山本衛, 森本康一, 日本再生医療学会総会(Web),   2019
  • 細胞低接着性コラーゲンの軟骨細胞分化誘導, 森本康一, 國井沙織, 堀内喜高, 伊田寛之, 平岡陽介, 山本衛, 日本再生医療学会総会(Web),   2019
  • 細胞低接着性コラーゲンを足場とした神経系細胞, 中野法彦, 兼清健志, 國井沙織, 森本康一, 尾前薫, 井出千束, 日本組織細胞化学会総会・学術集会講演プログラム・予稿集,   2018 09 29
  • 細胞低接着性コラーゲン(low adhesive scaffold collagen:LASCol)を用いた脊椎椎間板再生の試み, 武岡由樹, 由留部崇, 森本康一, 國井沙織, 深瀬直政, 竹森俊幸, 垣内裕司, 角谷賢一朗, 高田徹, 黒田良祐, 西田康太郎, 日本整形外科学会雑誌,   2018 08 30
  • 新規低接着性scaffold collagen(LASCol)はラット骨欠損モデルにおいて骨癒合を促進する, 竹森俊幸, 深瀬直政, 森本康一, 國井沙織, 由留部崇, 武岡由樹, 河本旭哉, 河本旭哉, 新倉隆宏, 原仁美, 秋末敏宏, 秋末敏宏, 黒田良祐, 日本整形外科学会雑誌,   2018 08 30
  • 細胞低接着性コラーゲン(Low Adhesive Scaffold Collagen:LASCol)による脊椎椎間板再生の可能性, 武岡由樹, 由留部崇, 國井沙織, 森本康一, 深瀬直政, 竹森俊幸, 垣内裕司, 伊藤雅明, 神田裕太郎, 角谷賢一朗, 高田徹, 黒田良祐, 西田康太郎, Journal of Spine Research,   2018 03 25
  • 細胞低接着性コラーゲン(Low Adhesive Scaffold Collagen:LASCol)による脊椎椎間板再生, 武岡由樹, 由留部崇, 國井沙織, 森本康一, 竹森俊幸, 深瀬直政, 垣内裕司, 角谷賢一朗, 黒田良祐, Japanese Journal of Rehabilitation Medicine,   2018
  • 細胞低接着性コラーゲンを足場とした神経細胞の挙動解析, 中野法彦, 兼清健志, 國井沙織, 森本康一, 尾前薫, 井出千束, 日本生化学会大会(Web),   2018
  • 真皮コラーゲンのLC‐MS/MSを用いた翻訳後修飾の解析, 永井宏平, 國井沙織, 中居由香理, 森本康一, 日本生化学会大会(Web),   2018
  • 新規低接着性コラーゲン(LASCol)はラット大腿骨骨欠損モデルにおいて骨癒合を促進する, 竹森俊幸, 深瀬直政, 森本康一, 國井沙織, 由留部崇, 武岡由樹, 原仁美, 河本旭哉, 河本旭哉, 黒田良祐, 秋末敏宏, 秋末敏宏, Japanese Journal of Rehabilitation Medicine,   2018
  • 骨組織から抽出したI型コラーゲンの骨芽細胞分化誘導, 國井沙織, 堀内喜高, 森本康一, 日本生化学会大会(Web),   2018
  • プロテアーゼによる骨組織の可溶化とタンパク質の同定, 國井沙織, 永井宏平, 森本康一, 日本生化学会大会(Web),   2018
  • 椎間板組織再生のための細胞低接着性コラーゲンゲルの開発, 森本康一, 國井沙織, 武岡由樹, 由留部崇, 加藤暢宏, 尾前薫, 黒田良祐, 日本バイオマテリアル学会大会予稿集(Web),   2018
  • 細胞低接着性コラーゲンによる神経細胞の活性化と組織再生機能の検証, 國井沙織, 兼清健志, 中野法彦, 加藤暢宏, 尾前薫, 井出千束, 森本康一, 日本バイオマテリアル学会大会予稿集(Web),   2018
  • 細胞低接着性コラーゲン培養により形成するES/iPS細胞の胚様体の解析, 森本康一, 國井沙織, 入江真純, 伊田寛之, 平岡陽介, 大貫喜嗣, 日本再生医療学会総会(Web),   2018
  • 細胞低接着性コラーゲン(LASCol)を用いた脊椎椎間板再生の可能性, 武岡由樹, 由留部崇, 國井沙織, 森本康一, 深瀬直政, 竹森俊幸, 垣内裕司, 辻本龍, 角谷賢一朗, 高田徹, 黒田良祐, 西田康太郎, 多血小板血漿(PRP)療法研究会プログラム・抄録集,   2018
  • 幹細胞から骨芽細胞への分化を短期間に誘導する細胞低接着性コラーゲン, 國井沙織, 堀内喜高, 伊田寛之, 平岡陽介, 森本康一, 日本再生医療学会総会(Web),   2018
  • 細胞低接着性コラーゲンを培養足場とした線維芽細胞スフェロイドの遺伝子発現量の変化, 森本康一, 國井沙織, 伊田寛之, 平岡陽介, 加藤暢宏, 日本バイオマテリアル学会大会予稿集(Web),   2017 11 13
  • 幹細胞から骨芽細胞への分化を短期間に誘導する細胞低接着性コラーゲン, 國井沙織, 堀内喜高, 山本衛, 伊田寛之, 平岡陽介, 森本康一, 日本バイオマテリアル学会大会予稿集(Web),   2017 11 13
  • 新奇な現象を生み出す“細胞低接着性コラーゲン”の培養足場としての活用, 森本康一, 國井沙織, 再生医療,   2017 02 01
  • 骨組織から精製したI型コラーゲンの骨芽細胞分化促進能, 國井沙織, 堀内喜高, 山本衛, 久保木芳徳, 森本康一, 日本骨代謝学会学術集会プログラム抄録集,   2017
  • 細胞低接着性コラーゲンで形成する線維芽細胞スフェロイドの解析, 國井沙織, 堀内喜高, 伊田寛之, 平岡陽介, 森本康一, 日本バイオマテリアル学会シンポジウム予稿集,   2016 11 21
  • マテリアルと細胞 細胞低接着性コラーゲンで形成する線維芽細胞スフェロイドの解析, 國井 沙織, 堀内 喜高, 伊田 寛之, 平岡 陽介, 森本 康一, 日本バイオマテリアル学会大会予稿集,   2016 11
  • 細胞低接着性コラーゲンが細胞増殖能に与える影響, 國井沙織, 加藤暢宏, 堀内喜高, 森本康一, 日本生化学会大会(Web),   2016 09
  • 細胞低接着性コラーゲンに起因する細胞の形態変化と分化誘導能の検証, 森本康一, DDS再生医療研究会プログラム・抄録集(Web),   2016
  • 低接着性コラーゲンが誘導する骨芽細胞への分化制御, 國井沙織, 山本衛, 堀内喜高, 赤星保光, 井田昌孝, 平岡陽介, 森本康一, 日本生化学会大会(Web),   2015 12
  • 線維芽細胞スフェロイドの運動能獲得性細胞足場の開発, 國井沙織, 堀内喜高, 井田昌孝, 平岡陽介, 森本康一, 日本バイオマテリアル学会大会予稿集,   2015 11 09
  • チタンとコラーゲンの反応について:インプラント蛋白発見に基づく展開, 久保木芳徳, 国井沙織, 森本康一, 古澤利武, 八上公利, 日本骨代謝学会学術集会プログラム抄録集,   2015 07
  • 低接着性コラーゲンを足場とした線維芽細胞の形態変化と運動能の獲得, KUNII SAORI, HORIUCHI YOSHITAKA, AKABOSHI YASUMITSU, KISHIGAMI SATOSHI, IDA MASATAKA, HIRAOKA YOSUKE, MORIMOTO KOICHI, 再生医療,   2015 02 01
  • in vivoで骨形成を誘導する低接着性コラーゲンの効果, MORIMOTO KOICHI, KUNII SAORI, HORIUCHI YOSHITAKA, AKABOSHI YASUMITSU, IDA MASATAKA, HIRAOKA YOSUKE, ITO HIROYUKI, YAMAMOTO EI, 再生医療,   2015 02 01
  • 潜在的な物性を引き出したコラーゲン・マテリアルの創製, 森本康一, 國井沙織, DDS再生医療研究会プログラム・抄録集(Web),   2015
  • 低接着性コラーゲンを足場とした線維芽細胞の挙動解析, 國井沙織, 堀内喜高, 井田昌孝, 平岡陽介, 森本康一, 日本生化学会大会(Web),   2015
  • 細胞とコラーゲンの接着には二面性がある―分子から細胞レベルまでの解析―, MORIMOTO KOICHI, 松本歯学,   2014 12 31
  • 細胞低接着性I型コラーゲンの開発と新生骨形成能の評価, MORIMOTO KOICHI, KUNII SAORI, YAMAMOTO EI, HORIUCHI YOSHITAKA, AKABOSHI YASUMITSU, ITO HIROYUKI, IDA MASATAKA, HIRAOKA YOSUKE, 日本バイオマテリアル学会大会予稿集,   2014 11 17
  • 細胞凝集能を有するI型コラーゲンの再生骨形成能の評価, KUNII SAORI, YAMAMOTO EI, MORIMOTO KOICHI, 日本骨代謝学会学術集会プログラム抄録集,   2014 07
  • in vitroでの線維芽細胞の凝集挙動の解析, KUNII SAORI, HIRAOKA YOSUKE, MORIMOTO KOICHI, 日本結合組織学会学術大会抄録集,   2014 06
  • 細胞凝集塊を誘導するコラーゲンの骨再生促進能の評価, KUNII SAORI, YAMAMOTO EI, ITO HIROYUKI, HIRAOKA YOSUKE, MORIMOTO KOICHI, 日本結合組織学会学術大会抄録集,   2014 06
  • カキ果実に含まれるキチナーゼの同定と精製, KUNII SAORI, KATAGIRI MINA, YAMANISHI HISAKO, OZAKI YOSHIHIKO, MORIMOTO KOICHI, 日本農芸化学会大会講演要旨集(Web),   2013 03 05
  • カキ果実に含まれるポリガラクツロン酸分解酵素阻害タンパク質の精製と阻害活性測定, 國井沙織, 片桐実菜, 山西妃早子, 尾崎嘉彦, 森本康一, 日本農芸化学会大会講演要旨集(Web),   2012 03 05
  • 黒毛和種牛の形質と影響タンパク質に対するSVMの検討, 堀貴裕, 中迫昇, 池上春香, 森本康一, 松本和也, 河本敬子, 電気関係学会関西連合大会講演論文集(CD-ROM),   2011 10 17
  • カキ果実の樹上成熟に伴うタンパク質発現量の変化と主要タンパク質の同定, 片桐実菜, 森本康一, 國井沙織, 阪井幸宏, 山西妃早子, 藤原真紀, 尾崎嘉彦, 日本食品科学工学会大会講演集,   2011 09 09
  • 黒毛和種とリムジン種の交雑家系(F2)のウシ僧帽筋のプロテオーム解析による経済形質バイオマーカーの探索, 武本淳史, 池上春香, 森本康一, 笹子奈々恵, 小林栄治, 松本和也, 日本畜産学会大会講演要旨,   2011 08 26
  • 糞を用いた組換え体タンパク質生産の可能性の検討, 孫谷 匡輝, 中村 文, 池上 春香, 奥山 紀之, 佐伯 和弘, 松本 和也, 若山 照彦, 森本 康一, 岸上 哲士, 細井 美彦, The Journal of Reproduction and Development,   2011 08 20
    Summary:【目的】トランスジェニック動物の乳汁や血液などを用いて,組換え体タンパク質を生産する研究が行われている。しかしながら,乳汁中からの産出は雌個体に限られ,生産量は産乳量などに依存する。血液中からの産出においても,個体への侵襲性などの問題がある。もし雌雄に関係なく排出される糞を用いた組換え体タンパク質の生産が可能ならば,産乳量や侵襲性などによる制限がなく有用物質の生産が期待できる。本研究では,糞を用いた組換え体タンパク質の生産の可能性を検討するため,緑色蛍光タンパク質(GFP)をモデルタンパク質とし,GFP遺伝子導入マウスの糞から蛍光を有するGFPの単離と特性解析を試みた。【方法】糞におけるGFPの局在を明らかにする為,樹脂切片において糞の蛍光観察を行った。次にリン酸バッファー中で糞を可溶および沈殿画分に分けSDS-PAGEを行い,Western Blot及び質量分析によりGFPの同定を行った。さらに,GFP遺伝子導入マウスの糞を用いて硫安分画,疎水クロマトグラフィー(Toyopearl Phenyl-650C)を行い,1M~0M硫安で溶出させ粗精製を行った。得られた各画分において,蛍光強度及びスペクトルを測定し,得られたGFPの特性解析を行った。【結果】GFP遺伝子導入マウスの糞は全体的に蛍光を有しており,特に糞表面において強い蛍光を示した。また糞中のGFPのほとんどが可溶画分に存在し,SDS-PAGEにおいてGFP遺伝子導入マウスの糞でのみ,GFPの大きさにあたる約27kDaでバンドが見られた。GFP抗体を用いたWestern Blotおよび質量分析によりこのタンパク質がGFPであることが明らかとなった。さらに,硫安分画及び疎水クロマトグラフィーで粗精製したタンパク質は強い蛍光を示し,GFP特有のスペクトルを有していた。以上の結果より,GFP遺伝子導入マウスの糞から,GFP特有の蛍光スペクトルと分子量をもつGFPが精製できる事が示され,糞を用いた組換え体タンパク質の生産の可能性が示唆された。
  • テロペプチド領域の切断で低下するI型コラーゲン会合体の熱安定性, 森本康一, 國井沙織, 外村辨一郎, 深田はるみ, 日本結合組織学会学術大会抄録集,   2011 05 18
  • テロペプチド領域を切除したコラーゲンにより変化する好中球の免疫ネットワークの解析, 國井沙織, 森本康一, 外村辨一郎, 日本結合組織学会学術大会抄録集,   2011 05 18
  • ウシの筋肉組織と脂肪組織のタンパク質発現比較, 池上 春香, 武本 淳史, 笹子 奈々恵, 森本 康一, 小林 栄治, 松本 和也, 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,   2010 12
  • ウシ僧帽筋に含まれるタンパク質の網羅的解析, 武本 淳史, 池上 春香, 笹子 奈々恵, 森本 康一, 小林 栄治, 松本 和也, 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,   2010 12
  • B205 Mechanical properties of callus formed in tibial defects in rats, SAKAKI Yusuke, KUNII Saori, MORIMOTO Koichi, YAMAMOTO Ei, Proceedings of the ... JSME Conference on Frontiers in Bioengineering,   2010 11 11
  • ラット脛骨欠損部位に形成される仮骨組織の力学的特性, 榊佑介, 國井沙織, 森本康一, 山本衛, 日本機械学会バイオフロンティア講演会講演論文集,   2010 11 11
  • テロペプチド領域の切断で低下するI型コラーゲン会合体の熱安定性, 國井沙織, 森本康一, 深田はるみ, 熱測定討論会講演要旨集,   2010 09 15
  • I型コラーゲンの線維形成能を制御するトリペプチド配列, 國井沙織, 永井宏平, 外村辨一郎, 森本康一, 日本結合組織学会学術大会抄録集,   2010 08 02
  • ウシ筋肉中タンパク質の網羅的プロテオーム解析, 武本淳史, 池上春香, 爲岡奈々恵, 森本康一, 小林栄治, 松本和也, 日本畜産学会大会講演要旨,   2010 03 28
  • ウシの枝肉形質に関与するバイオマーカー候補タンパク質のパスウェイ解析, 松本和也, 池上春香, 松橋珠子, 大谷健, 小林直彦, 森本康一, 日本畜産学会大会講演要旨,   2010 03 28
  • ウシ血清中に含まれるタンパク質の網羅的解析方法の確立, 池上春香, 松橋珠子, 小林直彦, 大谷健, 森本康一, 松本和也, 日本畜産学会大会講演要旨,   2010 03 28
  • 黒毛和種牛の形質と影響タンパク質に対する多変量解析の検討, 水野陽介, 河本敬子, 池上春香, 森本康一, 松本和也, 情報処理学会全国大会講演論文集,   2010 03 08
  • カキ果実Polygalacturonase‐inhibiting proteinの精製と阻害活性の解析, 森本康一, 國井沙織, 阪井幸宏, 中内道世, 尾崎嘉彦, 日本農芸化学会関西支部講演会講演要旨集,   2009 10 30
  • 酵素処理コラーゲンの末端配列決定と高次構造解析, 國井沙織, 永井宏平, 齋藤卓也, 外村辨一郎, 森本康一, 日本農芸化学会関西支部講演会講演要旨集,   2009 10 30
  • ウシの経済形質に関与する新規バイオマーカーAnnexin A5アイソフォームの同定, 池上春香, 園陽平, 永井宏平, 吉廣卓哉, 井上悦子, 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,   2009 09 28
  • アクチニダイン処理コラーゲンが誘導するNIH/3T3線維芽細胞の特異的な形態変化と運動能, 國井沙織, 森本康一, 生化学,   2009 09 25
  • カキ果実(Diospyros kaki Thunb.)に存在するPGIP(Polygalacturonase‐inhibiting protein)の解析, 阪井幸宏, 中内道世, 尾崎嘉彦, 森本康一, 日本農芸化学会大会講演要旨集,   2009 03 05
  • アクチニダイン処理コラーゲンが及ぼすNIH‐3T3細胞の形態変化, 國井沙織, 森本康一, 日本農芸化学会大会講演要旨集,   2009 03 05
  • MALDI‐MS/MSによる酵素処理ニワトリI型コラーゲンのC末端解析, 森本康一, 永井宏平, 國井沙織, 日本農芸化学会大会講演要旨集,   2009 03 05
  • Proteomic analysis of laminarialean algae under high temperature condition using 2-DE and homology-based protein identification., Nagai Kouhei, Ikegami Haruka, Kimura Hajime, Yotsukura Norishige, Morimoto Koichi, Abstracts for Annual Meeting of Japanese Proteomics Society,   2009
    Summary:[背景と目的] 近年の地球温暖化により、多くの生物の勢力地図が変わろうとしている。日本沿岸では各地で磯焼けにともなう種多様性の消失が進んでおり、温暖化対策が急務である。そのために我々は、高温耐性をもつコンブ目海藻の分子機構をプロテオーム解析により解明することを目的とした。またコンブ目海藻は多くがゲノム未解読であり、そのような生物のプロテオーム解析のモデルケースとしての手法確立も目指した。特に本発表では、高温ストレスで発現量が変化するタンパク質の同定を試みたので報告する。
    [材料と方法] コンブ目海藻カジメ(Ecklonia cava)とクロメ(Ecklonia kurome)の葉状部に含まれるタンパク質をエタノール/フェノール法により抽出し、二次元電気泳動(pI4-7、10% polyacrylamide gel、タンパク質量 200 μg)により分離した(文献1)。タンパク質115スポットをゲル内トリプシン消化し、MALDI-TOF/TOF型質量分析計によりMSと MS/MSスペクトルを得た。MS/MS スペクトルからアミノ酸配列を推定し(De novo sequence)、その推定配列と相同性をもつタンパク質をMS Blastにより検索した。さらに、10°C,20°C,30°Cで培養したカジメ個体のタンパク質を抽出して二次元電気泳動ゲルに展開した。各処理区でタンパク質発現量を定量比較し、増減のあったタンパク質を同様に決定した。
    [結果] 画像解析により、各二次元電気泳動ゲルから約700スポットをCBB染色により検出した。115スポットのタンパク質をMascot検索とMS Blast検索した結果、96スポット(84%)のタンパク質を同定することに成功した。高水温で誘導されるタンパク質はHSP70, HSP90, vanadium dependent bromoperoxidase (vBPO)などで、その発現量は増加していた。よって、コンブ目海藻にも高等植物と同様のHSPファミリータンパク質による高温耐性機構が考えられ、さらにvBPOなどの海藻特異的なタンパク質が強く関与する特異な高温耐性メカニズムを有する可能性が示唆された。
    文献 1) K. Nagai, et al., Electrophoresis 29, 672-681 (2008)
  • Discovery of biomarker candidates, Annexin A5 isoforms, related to bovine economic traits by large-scale proteomic data analysis, Ikegami Haruka, Morimoto Koichi, Matsumoto Kazuya, Sono Youhei, Nagai Kouhei, Yoshihiro Takuya, Inoue Etsuko, Kobayashi Naohiko, Matsuhashi Tanako, Ohtani Tsuyoshi, Nakagawa Masaru, Abstracts for Annual Meeting of Japanese Proteomics Society,   2009
    Summary:背景 : 近年、農学分野においてもプロテオミクス研究が進展している一方、家畜の育種・生産分野では、家畜の育種選抜・生産や病気を診断するバイオマーカーの探索はほとんど実施されていない。我々は、ウシの経済形質(枝肉重量,ロース芯面積,BMS No.(脂肪交雑基準)など)に関与するバイオマーカーの探索を目的として、10944頭のウシの生物情報(血統や枝肉形質43項目)をデータベース化するとともに、ウシ白色脂肪組織を用いた大規模プロテオーム解析システムを構築した(文献[1])。
    方法 :データベースに登録したウシから任意に220個体を抽出し、白色脂肪組織に発現しているタンパク質について、二次元電気泳動を用いた発現量解析とMALDI-TOF/TOF型質量分析機(4700, ABI)を用いたタンパク質の同定をおこなった。プロテオーム解析データはLIMSソフトウェアBIOPRISM(NEC)で一元管理し、ウシの生物情報43項目とプロテオーム解析情報(879個のタンパク質発現量)を用いて統計解析をおこなった。
    結果 : ウシ白色脂肪組織のタンパク質から、詳細なMS/MSスペクトルの解析により2種類のAnnexin A5アイソフォーム(CaBP33, CaBP37)を検出・同定した。CaBP33とCaBP37両方を発現する個体群(96頭),CaBP33のみを発現する個体群(106頭),CaBP37のみを発現する個体群(18頭)に分類した。各群間の生物情報を比較したところ、月齢には優位な差が見られないのに対し、枝肉重量とロース芯(最長胸筋)面積について差が見られ、CaBP33とCaBP37両方を発現する個体群がどちらか一方を発現する個体群よりも小さい傾向にあった。また、分類した各群間のAnnexin A5以外のタンパク質発現量を比較したところ、28スポットのタンパク質発現量に差が見られた(p<0.05)。さらに、200検体のAnnexin A5発現パターンで分類しない個体群から枝肉重量の大きい個体群(>平均値+標準偏差の個体,n=27)と小さい個体群(<平均値-標準偏差の個体,n=26)を抽出し、各タンパク質の発現量を比較したところ、44スポットについて有意差があり(P<0.05)、5スポット2種類のタンパク質についてAnnexin A5アイソフォーム発現パターンで分類した群間で発現量に差のあったタンパク質と共通していた。各タンパク質のバイオマーカーとしての検証をおこなうとともに、経済形質の予測診断に最適なタンパク質バイオマーカーの組み合わせに関して検討している。
    文献[1] K. Nagai et al., Animal Sci. J. Vol.79, No.4, 2008
  • Examination of Effective Culture Methods for Rabbit Preantral Follicles, Sugimoto Hironobu, Miyamoto Yuki, Tsuji Yoko, Morimoto Kouichi, Taniguchi Takeshi, Morimoto Yoshiharu, Hosoi Yoshihiko, Journal of Mammalian Ova Research,   2009
    Summary:To determine an effective in vitro culture method of rabbit immature ovarian follicles, preantral follicles (200 to 299 μm in diameter) were cultured by the whole follicle culture method and by the open type culture method. The use of tuna matrix collagen (MC) as a medium material to maintain 3-dimensional structure of the follicles was also investigated for the open type culture method. In the whole follicle culture method, the diameter of the preantral follicles increased from 252.8 ± 2.7 μm to 395.6 ± 9.6 μm. However, culture for oocyte maturation was not possible because most of the oocytes had degenerated. In the open culture method, in which oocyte-granulosa cell complexes (OGs) collected from preantral follicles were cultured, the rates of the oocytes that resumed meiosis were 1.6%, 5.4%, and 73.7% in 0, 0.3, and 3 mg/ml of MC supplementation, respectively. Morphologically, the addition of MC was found to show a beneficial effect for maintenance of the three-dimensional structure of the follicles. In conclusion, open type culture with MC supplementation seems to be a most appropriate culture method for rabbit immature follicles.
  • プロテオーム解析を用いたウメ環境ストレス耐性個体選抜マーカーの作成, 花田裕美, 橋本広祐, 根来圭一, 林恭平, 永井宏平, 池上春香, 森本康一, 園芸学研究 別冊,   2008 09 27
  • アコヤガイ貝殻タンパク質の同定, 矢野昌人, 永井宏平, 森本康一, 宮本裕史, 日本動物学会大会予稿集,   2008 08 20
  • アコヤガイ真珠層タンパク質の解析, 矢野昌人, 永井宏平, 森本康一, 宮本裕史, マリンバイオテクノロジー学会大会講演要旨集,   2008 05 24
  • 黒毛和牛の肉質に関連するバイオマーカーの探索, 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,   2008 03 27
  • ブランド牛(飛騨牛)の繁殖農家のための種雄牛選択支援システム, SHI Linjing, 井上悦子, 吉廣卓哉, 永井宏平, 池上春香, 松橋珠子, 小林直彦, 森本康一, 松本和也, 中川優, 情報処理学会全国大会講演論文集,   2008 03 13
  • コラーゲンにより誘導される好中球活性化機構の解析, 國井沙織, 柴野三智子, 斎藤卓也, 森本康一, 日本農芸化学会大会講演要旨集,   2008 03 05
  • 二青ウメの葉において乾燥ストレス処理で発現が誘導されるASR遺伝子の解析, 橋本広祐, 永井宏平, 池上春香, WONGCHAOCHANT Shermarl, 林恭平, 根来圭一, 森本康一, 花田裕美, 日本農芸化学会大会講演要旨集,   2008 03 05
  • Amide‐80とODS‐100Vを用いたペプチドマッピング, 永井宏平, 佐藤真治, 森山弘之, 森本康一, 日本農芸化学会大会講演要旨集,   2008 03 05
  • コンブ目海藻カジメにおいて水温依存的に発現するタンパク質の同定, 永井宏平, 四ツ倉典滋, 加藤弘美, 田中俊充, 木村創, 森本康一, 日本農芸化学会大会講演要旨集,   2008 03 05
  • Large-scale proteomic analysis of bovine white adipose tissue - Discovery of protein biomarker candidates related to carcass and meat quality traits in Japanese Black beef cattle (Wagyu) -, Ikegami Haruka, Morimoto Koichi, Matsumoto Kazuya, Sono Youhei, Nagai Kouhei, Yoshihiro Takuya, Inoue Etsuko, Kobayashi Naohiko, Matsuhashi Tamako, Otani Tsuyoshi, Nakagawa Masaru, Abstracts for Annual Meeting of Japanese Proteomics Society,   2008
    Summary:Introduction: In livestock researches, the proteomics has the potential to be a method for identification of biomarker related to carcass and meat quality traits. Two-dimensional gel electrophoresis (2-DE) has been used in combination with MS identification of specific proteins as the proteomics tool. However, proteins that contribute to individual variations in carcass and meat quality traits are not fully understood. In order to discover protein biomarkers involved in carcass and meat quality traits in Japanese Black beef cattle, we have advanced large-scale proteomic analysis of bovine white adipose tissue (WAT), using 2-DE and mass spectrometry.
    Methods:Proteins extracted from the WAT were separated by 2-DE and visualized using SYPRO Ruby. The separated proteins were identified by MALDI-TOF/TOF and Mascot search engine (Matrix science). Expression levels of the separated proteins were calculated with TT900 and Progenesis PG220 software (Nonlinear dynamics). Finally, correlation of protein expression and the carcass trait was examined by the statistical analysis using the quantitative expression values of each protein in the WAT from 133 animals.
    Results:In our study, 879 protein spots were detected on the 2-DE gels. Of these, a total of 459 protein spots were extensively identified by MALDI-TOF/TOF. As a result of statistical analysis, it was shown that in higher carcass weight (CW) group, 95 protein spots were up-regulated and two protein spots were down-regulated compared with those in lower CW group. These proteins identified as biomarker candidates to the carcass weight in Japanese Black beef cattle are involved in a variety of functions, including energy metabolism, cell structure, cell defense, transport, and signal transduction.
    Acknowledgement:This work was supported by a grant from the Wakayama Prefecture Collaboration of Regional Entities for the Advancement of Technological Excellence of JST.
  • Proteomic analysis of transgenic mice bearing Spinacia omega-6 desaturase gene, Sono Youhei, Ikegami Haruka, Nagai Kouhei, Taguchi Yoshitomo, Hosoi Yoshihiko, Saeki Kazuhiro, Iritani Akira, Morimoto Koichi, Matsumoto kazuya, Abstracts for Annual Meeting of Japanese Proteomics Society,   2008
    Summary:Introduction: We observed that two-lines of transgenic mice ubiquitously expressing Spinacia microsomal fad2 gene for omega-6 desaturase were lean phenotype. These mice were shown to be resistant to obesity induced by high fat diet, indicating their basal metabolic rate is also elevated. Here, we investigated about differentially expressed proteins between two-lines of transgenic and wild-type mice.
    Methods: Two mg of white adipose tissue and 1mg of skeletal muscle were homogenized and added 4uL of lysis buffer, containing 7M urea, 2M thiourea, 4% CHAPS, 0.5%IPG buffer pH 3-11 nonlinear,0.05%TBP and protease inhibitor cocktail. Proteins were separated by 2-DE and were visualized using SYPRO Ruby. The separated proteins were identified by MALDI-TOF/TOF and Mascot search (Matrix science). Expression levels of the separated proteins calculated with PG220 and TT900 software (Nonlinear dynamics). The statistical analysis of quantitative expression values of each protein was done between two lines of transgenic and wild type mice.
    Result: In the white adipose tissue and skeletal muscle, 810 and 635 protein spots were commonly detected on 2-DE gels and 329 and 315 protein spots were identified by MALDI-TOF/TOF, respectively. When two lines of transgenic and wild-type mice were compared in the white adipose tissue, we observed that 38 protein spots were differentially expressed. These included glycolysis-related proteins. Moreover, 30 proteins spots were shown to be differential expressed in comparison between skeletal muscle from two lines of transgenic and wild-type mice. These proteins were involving in glycolysis and mitochondrial function. The current data suggest that up-regulated energy metabolism may induce lean phenotype in transgenic mice bearing fad2 gene.
    Acknowledgement: This work was supported by grant from the Wakayama Prefecture Collaboration of Regional Entities for the Advancement of Technological Excellence of JST.
  • 乾燥ストレスによりウメで特異発現をするASR(Abscisic acid stress ripening like protein)タンパク質の同定, 花田裕美, 橋本広祐, 根来圭一, 林恭平, 永井宏平, 池上春香, 森本康一, 生化学,   2008
  • 酵素処理コラーゲンが誘導するマウス好中球の機能発現, 國井沙織, 森本康一, 生化学,   2008
  • 乾燥ストレスによりウメで特異発現をするASR(Abscisic acid stress ripening like protein)タンパク質の同定, 花田裕美, 橋本広祐, 根来圭一, 林恭平, 永井宏平, 池上春香, 森本康一, 質量分析総合討論会講演要旨集,   2008
  • ウシの枝肉形質を規定するバイオマーカーの探索―質量分析を用いたタンパク質アイソフォームの解析―, 池上春香, 永井宏平, 吉廣卓哉, 井上悦子, 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, 質量分析総合討論会講演要旨集,   2008
  • ホウレンソウ由来Δ12脂肪酸不飽和化酵素遺伝子導入マウスのプロテオーム解析, 園陽平, 池上春香, 永井宏平, 田口善智, 細井美彦, 佐伯和弘, 入谷明, 森本康一, 松本和也, 生化学,   2008
  • バイオミネラリゼーションにおけるプロテオーム解析の応用, 矢野昌人, 永井宏平, 森本康一, 宮本裕史, 質量分析総合討論会講演要旨集,   2008
  • 黒毛和牛の枝肉形質を診断するバイオマーカー候補タンパク質の探索:白色脂肪組織の大規模プロテオーム解析, 池上春香, 園陽平, 永井宏平, 吉廣卓哉, 井上悦子, 小林直彦, 松橋珠子, 大谷健, 中川優, 森本康一, 松本和也, 生化学,   2008
  • 酵素処理コラーゲン培養が及ぼす前骨髄性白血病細胞HL60の挙動変化, 國井沙織, 森本康一, 生化学,   2008
  • コンブ目海藻を対象としたプロテオーム解析~ゲノム解読されていない生物へのアプローチ~, 永井宏平, 四ツ倉典滋, 池上春香, 木村創, 森本康一, 質量分析総合討論会講演要旨集,   2008
  • ゲノム未解読の農産物で特異的に発現する機能性タンパク質のプロテオーム解析, 森本康一, 質量分析総合討論会講演要旨集,   2008
  • 3P049 Intermolecular Propagation of βrich Protofibril(Proteins-stability, folding, and other physicochemical properties,Poster Presentations), Hayashi Itsuho, Nakatsuka Ken, Maeno Akihiro, Morimoto Koichi, Akasaka Kazuyuki, Biophysics,   2007 11 20
  • 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,   2007 09 26
    Summary:【背景】
    和歌山県は全国シェア第1位のウメとカキの他にも,マダイ稚魚や海藻など地域の特性を活かした優良な農業資源が豊富である.また,特産優良和牛である熊野牛のブランド化を推進している.私どもは科学的根拠に基づく手段として優良和牛を検定するため,生物学的データ量が蓄積されている"飛騨牛"をモデル畜産動物とし,その血統・肉質情報をデータベース化し,白色脂肪組織に発現している全タンパク質の網羅的同定を進めている.本発表では,これまでに報告のないプロテオーム解析手法を用いて有用形質と相関を示すタンパク質バイオマーカーを探索したので報告する.
    【材料と方法】
    飛騨牛の白色脂肪組織 1 gに対して抽出溶液(7M Urea, 2M Thiourea, 4% CHAPS, 0.5% IPG buffer, 0.05% TBP, Protease inhibitor )1 mlを加えてホモジナイズし,遠心分離により沈殿を除去した.タンパク質150 ugを二次元電気泳動にて分離し,SyproRubyを用いて染色,ゲルイメージを得た.分離したタンパク質スポットをトリプシンにより切断し,MALDI-TOF/TOF型質量分析計(4700 Proteomic Analyzer,ABI)を用いてMSとMS/MS測定した.得られたスペクトルデータを用いてPMFおよびMS/MS IonseearchでMASCOT検索し,タンパク質を同定した.取得したゲルイメージはTT900とProgenesis220(nonlinear dynamics)を用いて画像解析し,得られた各タンパク質スポットの定量値を各形質データ(個体データ,血統データなど7項目,肉質データ22項目)に対してプロットし,タンパク質発現と各生物情報との相関関係を調べた.
    【結果】
    脂肪組織1gから平均して3.5mgのタンパク質が回収された.得られたウシ121検体分のゲルイメージの画像解析から,個体依存的に発現する特異なタンパク質(A)を同定した.Aの発現パターンと各形質の相関を調べた結果,肉質データ1項目について統計的に有意な相関が認められた.現在,解析個体数を増やすとともに,Aと肉質との関係について生化学的な考察を含めた統合解析を進めている.
    尚,本研究はJST和歌山県地域結集型共同研究事業費で行なった.
  • Microbiological Evaluation of Fruits Certified as Specially Grown Agricultural Products by an Accredited Certification Agency, Izumi Hidemi, Morimoto Koichi, Yamawaki Nobuyuki, Murakami Yukari, Kida Kaori, Memoirs of the School of Biology-Oriented Science and Technology of Kinki University,   2007 09 01
    Summary:The partial organic fruits (Japanese apricot, Japanese plum, persimmon, satsuma mandarin, and lemon) grown under restricted inorganic practices and certified as Specially Grown Agricultural products by a Wakayama Prefecture-accredited certification agency were evaluated for microbial quality and safety in 2004 to 2006. Microbial population in most whole fruit or peel and flesh of fruit was below the detection level (2.4log CFU/g for bacteria and 3.0log CFU/g for fungi). Among all samples, the peel of satsuma mandarin showed the highest counts of 3.9log CFU/g for total bacteria in 2004 and 5.0log CFU/g for fungi in 2005. The microbial counts in the flesh of all fruits were below the detection level and the pH values of flesh of all fruits were as low as 3.2 except for persimmon. Molds comprised approximately 80% of the total isolates in all fruits. The most frequently isolated mold genera were Alternaria, Diaporthe, Pestalotia, Phialophora, and Phanerochaete, which were phytopathogenic organisms. No human pathogens such as Salmonella and Escherichia coli O157:H7 were detected from any of the samples. These results suggest that the partial organic fruits do not have high microbial risk due to the farming methods used.
  • アコヤガイ貝殻に存在する新規tyrosinase, 矢野昌人, 永井宏平, 森本康一, 宮本裕史, マリンバイオテクノロジー学会大会講演要旨集,   2007 05 26
  • 酵素分解コラーゲンが引き起こす好中球の形態変化と運動能獲得の解析, 國井沙織, 柴野三智子, 斎藤卓也, 森本康一, 日本農芸化学会大会講演要旨集,   2007 03 05
  • 酵素処理コラーゲンの熱安定性と構造特性, 森本康一, 國井沙織, 柴野三智子, 斎藤卓也, 日本農芸化学会大会講演要旨集,   2007 03 05
  • Peptide mapping using a normal phase Amide-80 column and a reversed phase ODS C-18 column, Nagai Kouhei, Morimoto Koichi, Sato Shinji, Moriyama Hiroyuki, Abstracts for Annual Meeting of Japanese Proteomics Society,   2007
    Summary:[目的]
    ODS C-18カラムを用いた逆相系HPLCは、高い分解能を示し、また、遊離液に不揮発性塩を必要としないことから質量分析 (LC-MS)の前段階として、広く用いられている。プロテオーム解析においても、トリプシン分解したタンパク質の断片の分離に、ODS C-18カラムが最も広く用いられている。しかし、全てのペプチド断片が、このカラムにより検出されることは非常に稀である。本研究では、順相系カラムAmide 80をODS C-18と併用して使用することで、LC-MS解析におけるタンパク質のSequence coverage (SC)を向上させることを目指した。

    [方法]
    ウシ血清アルブミン(BSA)を還元カルボキシメチル化した後、溶液中でトリプシン消化した。得られたBSA分解物をODS C-18カラム (0.3 mm ID×100 mm)、もしくはAmide 80カラム (0.3 mm ID×100 mm) を用いたHPLCにて分離し、Linear Ion trap型質量分析器Q-Trap (ABI)によってMS/MS解析した。得られたデータをMASCOT検索にかけ、BSAのSCを評価した。

    [結果]
    10 pmol のBSA 分解物のLC-MS/MS解析を行い、MASCOT検索にかけた。ODS C-18カラムを用いた場合、58-60本のBSAのピークが検出され、そのSCは67.7%と求められた。Amide 80では、49-50本のピークが検出され、SCは50% と求められた。ODS C-18では親水性の高いペプチドが検出されやすく、Amide 80 では疎水性の高いペプチドが検出されやすい傾向がみられた。このため、両カラムを併用することでSC を79.1%まで向上させることができた。本方法は、アイソフォームの同定やタンパク質修飾の解析にも応用可能であると考えられる。
  • 好中球運動能の新規亢進因子として機能するアクチニダイン処理I型コラーゲン, 國井沙織, 柴野三智子, 堀内喜高, 齋藤卓也, 森本康一, 日本結合組織学会学術大会抄録集,   2007
  • アクチニダイン処理コラーゲンの高次構造が誘導する好中球の細胞動態と運動能, 國井沙織, 柴野三智子, 堀内喜高, 齋藤卓也, 森本康一, 生化学,   2007
  • アクチニダイン処理コラーゲンにより誘導される好中球の新奇な運動能, 森本康一, 國井沙織, 柴野三智子, 齋藤卓也, 日本結合組織学会学術大会抄録集,   2007
  • 優良肉質を規定するバイオマーカー候補の探索:飛騨牛をモデルとして, 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, 生化学,   2007
  • アコヤガイ貝殻に存在する新規tyrosinase, 矢野昌人, 永井宏平, 森本康一, 宮本裕史, 生化学,   2007
  • コンブ目海藻カジメのタンパク質発現に与える水温の影響, 永井宏平, 加藤弘美, 四ツ倉典滋, 田中俊充, 木村創, 森本康一, 生化学,   2007
  • 酵素処理で得られた低線維形成能コラーゲン会合体の熱安定性, 國井沙織, 柴野三智子, 齋藤卓也, 外村辨一郎, 森本康一, 日本農芸化学会関西支部講演会講演要旨集,   2007
  • Thermal stability and structural feature of actinidain-hydrolyzed collagen self-assembly, Meeting Program of EABS & BSJ,   2006 12 , Meeting Program of EABS & BSJ
  • SEM observation of neutrophil motility in collagen matrix hydrolyzed by actinidain peptidase,   2006 09
  • Research on proteome analysis in mouse ovary, Uenaka Takahiro, Nagai Kouhei, Ikegami Haruka, Matsumoto Kazuya, Morimoto Kouichi, Saeki Kazuhiro, Hosoi Yoshihiko, Iritani Akira, Abstracts for Annual Meeting of Japanese Proteomics Society,   2006 07
    Summary:卵巣の機能は脳下垂体から分泌される2つのホルモンによって制御されている。ひとつは卵胞刺激ホルモン(FSH:follicle-stimulating hormone)で、これは卵子を育む卵胞の発達を促進する。その後、FSHにより十分に発達した卵胞に作用するもうひとつのホルモン、黄体形成ホルモン(LH:luteinzing hormone)が卵巣に届き、卵胞からの排卵が誘起される。本実験では、FSH様作用を持つ妊馬血清性腺刺激ホルモン(PMSG:Pregnant mare serum gonadotropin)とLH様作用を持つヒト絨毛性性腺刺激ホルモン(hCG:Human chorionic gonadotropin)の投与によるPMSG-hCG法を用いて実験動物であるマウスの卵巣を調整した。卵巣の解析ではこれまでに、ホルモンレセプターやイオンチャネルなど特定のタンパク質に注目した研究がなされてきた。また、排卵中の変化を網羅的に研究した例では、組織の質量を測定した報告がある。しかし卵巣組織に起こる現象を発現しているタンパク質全体の変化から、網羅的に研究した例はない。本研究では、卵巣に含まれる全タンパク質の抽出法、二次元電気泳動法、およびタンデム質料分析器を用いた網羅的タンパク質の同定法を試みた。まず、ICR系マウスから処理前区、PMSG投与後24時間区、PMSG投与後48時間区、hCG投与後20時間区の卵巣をサンプルとした。卵巣で発現しているタンパク質を網羅的に解析することを目的にpI3-11、10%アクリルアミドゲルを用いて二次元電気泳動を行った。その結果1400以上のタンパク質を分離することに成功し、そのうち256スポットにおいてMOLDI TOF-MSを用い、解析・同定を行った。その結果、transferrinや、glutathion S-transferase、heat shock proteinなど生体で多く発現しているタンパク質の192スポットを同定した。さらに、各区画の卵巣から得たタンパク質抽出液を、二次元電気泳動を行い経時的な発現量の変化を測定した。その結果、ホルモン投与後に発現量の上がるtransketolaseやhCG投与後20時間後に発現量が上昇するvimentin、またホルモンの投与後に発現量が減少する14-3-3 proteinなど様々な挙動を示すタンパク質を解析した。また処理により解糖系クエン酸回路に寄与する酵素の発現が一様に上昇した。以上の結果から性腺刺激ホルモンの感作を受けた卵巣では排卵までの過程で解糖系のタンパク質の発現量が亢進していることが示された。尚、本研究はJST和歌山県地域結集型共同研究費で行われた。
  • Proteome analysis using leaf tissues of Japanese apricot (Prunus mume) at drought or salt stress conditions, 20th IUBMB international congress of Biochemistry and Molecular Biology and 11th FAOBMB congress,   2006 06 , 20th IUBMB international congress of Biochemistry and Molecular Biology and 11th FAOBMB congress
  • Comparison of proteomic profiles between several species of Japan kelp using 2D-PAGE and MALDI-TOF/TOF, 20th IUBMB international congress of Biochemistry and Molecular Biology and 11th FAOBMB congress,   2006 06 , 20th IUBMB international congress of Biochemistry and Molecular Biology and 11th FAOBMB congress
  • Purification and characterization of thaumatin-like protein of kiwifruit, 20th IUBMB international congress of Biochemistry and Molecular Biology and 11th FAOBMB congress,   2006 06 , 20th IUBMB international congress of Biochemistry and Molecular Biology and 11th FAOBMB congress
  • Neutrophil motility induced by actinidain-hydrolyzed collagen matrix, 20th IUBMB international congress of Biochemistry and Molecular Biology and 11th FAOBMB congress,   2006 06 , 20th IUBMB international congress of Biochemistry and Molecular Biology and 11th FAOBMB congress
  • Specific binding of thioflavin T to collagen fibril lacking beta-form structure, 20th IUBMB international congress of Biochemistry and Molecular Biology and 11th FAOBMB congress,   2006 06 , 20th IUBMB international congress of Biochemistry and Molecular Biology and 11th FAOBMB congress
  • 乾燥および塩ストレス処理にウメ葉組織で発現するプロテオームの解析, 花田裕美, 根来圭一, 永井宏平, 池上春香, 森本康一, 園芸学会雑誌 別冊,   2006 03 29
  • キウイフルーツに含まれるThaumatin‐like proteinの精製と構造解析, 森本康一, 天尾雅, 池上春香, 串田武司, 森山弘之, 井上國世, 日本農芸化学会大会講演要旨集,   2006 03 05
  • 二次元電気泳動とMALDI‐TOF/TOFを用いた暖海性コンブ目海藻のプロテオーム情報の種間比較, 池上春香, 森本康一, 永井宏平, 四ツ倉典滋, 木村創, 日本農芸化学会大会講演要旨集,   2006 03 05
  • アクチニダン処理コラーゲン培養基質で生じる好中球の特異的挙動, 國井沙織, 柴野三智子, 斎藤卓也, 森本康一, 日本農芸化学会大会講演要旨集,   2006 03 05
  • 乾燥,塩ストレスに対するウメの葉を用いたプロテオーム解析, 花田裕美, 根来圭一, 永井宏平, 池上春香, 森本康一, 日本農芸化学会大会講演要旨集,   2006 03 05
  • MALDI‐TOF/TOFによる相同性を利用した暖海性コンブ目海藻クロメのタンパク質の同定, 永井宏平, 池上春香, 森本康一, 木村創, 四ツ倉典滋, 日本農芸化学会大会講演要旨集,   2006 03 05
  • 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, 森本康一, 佐伯和弘, 細井美彦, 入谷明, 日本分子生物学会年会講演要旨集,   2005 11 25
  • アコヤガイ貝殻有機マトリックスタンパク質の同定, 矢野昌人, 永井宏平, 森本康一, 宮本裕史, 日本分子生物学会年会講演要旨集,   2005 11 25
  • ウメ(Prunus mume)の葉組織において乾燥および塩ストレス処理により特異的に発現するタンパク質のプロテオーム解析, 花田裕美, 根来圭一, 永井宏平, 池上春香, 森本康一, 日本分子生物学会年会講演要旨集,   2005 11 25
  • 飛騨牛白色脂肪組織のプロテオーム解析, 池上春香, 松本和也, 森本康一, 永井宏平, 上中崇裕, SHIN Sunuku, 小林直彦, 大谷健, 入谷明, 日本分子生物学会年会講演要旨集,   2005 11 25
  • 褐藻(Ecklonia kurome)から抽出した蛋白質の2-DE MALDI-MS/MS解析(原標題は英語), 森本康一, 永井宏平, 池上春香, 木村創, 四ツ倉典滋, 日本分子生物学会年会講演要旨集,   2005 11 25
  • 2P044 Evidence for nanostructural and superstructural differences of peptidase-treated collagen matrices, Morimoto K, Kunii S, Shibano M, Saito T, Biophysics,   2005 10 19
  • 2P237 Microscopic analysis of neutrophil cell mobility in novel 3-D network collagen matrix lacking telopeptides, Kunii S, Saito T, Shibano M, Morimoto K, Biophysics,   2005 10 19
  • Ultra-structural findings of type I collagen matrix generated by actinidain limited-hydrolysis,   2005 10
  • Proteomic analysis of brown alga, Ecklonia kurome obtained at Wakayama coast by MALDI-TOF/TOF,   2005 10
  • Proteomic analysis of Prunus mume leaves, HANADA H, NAGAI K, IKEGAMI H, MORIMOTO K, 園芸学会雑誌. 別冊, 園芸学会大会研究発表,   2005 10 01
  • 飛騨牛白色脂肪組織のプロテオーム解析, 池上春香, 松本和也, 森本康一, 永井宏平, 上中崇裕, 申承旭, 小林直彦, 大谷健, 入谷明, 日本畜産学会大会講演要旨,   2005 08 25
  • LIMSを用いた生物資源の統合的なプロテオーム解析, 永井宏平, 森本康一, 吉広卓哉, 池上春香, 剣持聡久, 上条憲一, 奥野充利, 中川優, 松本和也, 日本畜産学会大会講演要旨,   2005 08 25
  • 微生物酵素を用いたカキ果実剥皮技術の開発, 阪井幸宏, 尾崎嘉彦, 中内道世, 森本康一, 坂井拓夫, 日本食品科学工学会大会講演集,   2005 08 20
  • Comparison of Biochemical Properties of Novel Actinidain‐hydrolyzed Collagen and Pepsin‐hydrolzyed Collagen, 国井沙織, 森本康一, 近畿大学先端技術総合研究所紀要,   2005 03 15
  • 高塩濃度によるアクチニダイン・ペプチダーゼ活性の回復, 森本康一, 広垣和洋, 橋本浩治, 井上国世, 日本農芸化学会大会講演要旨集,   2005 03 05
  • クロマトフォーカシングHPLCを用いたアクチニダイン・アイソザイムの分離, 串田武司, 池上春香, 永井宏平, 三苫恵民, 井上国世, 森本康一, 日本農芸化学会大会講演要旨集,   2005 03 05
  • コンブ目海藻のプロテオーム解析に向けた二次元電気泳動法の確立, 永井宏平, 池上春香, 木村創, 四ツ倉典滋, 森本康一, 日本農芸化学会大会講演要旨集,   2005 03 05
  • 異なる動物種から調製したアクチニダイン処理I型コラーゲンの生化学的性質, 国井沙織, 斎藤卓也, 森本康一, 日本農芸化学会大会講演要旨集,   2005 03 05
  • ウメ(Prunus mume)の品種特異的発現タンパク質のプロテオーム解析, 花田裕美, 永井宏平, 池上春香, 森本康一, 日本農芸化学会大会講演要旨集,   2005 03 05
  • Cell-free expression of prion protein and the structural study using high pressure fluorescence spectroscopy,   2004 12
  • Interaction of Thioflavin T fluorescence dye with collagen fibrils,   2004 12
  • キハダマグロ由来I型コラーゲンの線維と網目会合体の分光学的解析, 川端和也, 浜野香織, 赤坂一之, 森本康一, 生化学,   2004 11 25
  • 2P026 Interaction of thioflavin T fluorescence dye with collagen fibrils, Morimoto K, Kawabata K, Kunii S, Akasaka K, Biophysics,   2004 11 10
  • 2P025 Cell-free exnression of prion protein and the structural study using high pressure fluorescence spectroscopy, Gaikwad, Jyoti U, Sasaki K, Komabayashi H, Morimoto K, Akasaka K, Biophysics,   2004 11 10
  • The hemostatic effectveness of tuna fish collagen,   2004 10
  • Spectroscopic and morphological analysis of atelocollagen fibril formation,   2004 09
  • Morphological stereo-observation of neutrophil adhesion onto chiken actinidain-processed atelocollagen by scanning electron microscopy,   2004 09
  • Three-dimensional observation of neutrophil and actinidain-processed atelocollagen by scanning electron microscopy, Matrix Biology Society of Australia and New Zealand,   2004 09 , Matrix Biology Society of Australia and New Zealand
  • Unique migration and accumulation activities of neutrophil within actinidain-processed atelocollagen by scanning electron microscopy, Matrix Biology of Australia and New Zealand,   2004 09 , Matrix Biology of Australia and New Zealand
  • 医療用魚コラーゲンの開発, 吉川隆章, 森本康一, 飯田仁, 奥村紀子, 野々村昭孝, 高倉義典, 日本整形外科学会雑誌,   2004 08 25
  • Morphological observation of actinidain-processed atelocollagen and neutrophil adhesion to the collagen by scanning electron microscopy, 8th Asia-Pacific conference on electron microscopy,   2004 06 , 8th Asia-Pacific conference on electron microscopy
  • Analysis of atelocollagen fibril formation by using thioflavin T, The 1st Pacific-Rim International Conference on Protein Science,   2004 04 , The 1st Pacific-Rim International Conference on Protein Science
  • Morphological analysis and hemostatic efficacy of actinidain-processed atelocollagen, The 1st Pacific-Rim International Conference on Protein Science,   2004 04 , The 1st Pacific-Rim International Conference on Protein Science
  • ハイドロキシアパタイトパターン上への蛋白質吸着, 楠正暢, 川島将実, 西川博昭, 本津茂樹, 森本康一, 川合知二, 応用物理学関係連合講演会講演予稿集,   2004 03 28
  • アクチニダインのエステラーゼ活性に対する塩の効果, 森本康一, 古田えりの, 橋本浩治, 井上国世, 日本農芸化学会大会講演要旨集,   2004 03 05
  • アクチニダイン処理アテロコラーゲンの止血効果, 国井沙織, 森本康一, 斉藤卓也, 吉川隆章, 外村弁一郎, 日本農芸化学会大会講演要旨集,   2004 03 05
  • アクチニダインのエステラーゼ活性に対するNaClとアルコールの影響, 森本康一, 古田えりの, 橋本浩治, 井上国世, 日本農芸化学会関西支部講演会講演要旨集,   2003 10 04
  • アクチニダイン処理マグロI型コラーゲンの血液凝固作用, 国井沙織, 森本康一, 吉川隆章, 外村弁一郎, 日本農芸化学会関西支部講演会講演要旨集,   2003 10 04
  • 電子顕微鏡観察によるアクチニダイン限定分解アテロコラーゲンの構造解析, 浜野香織, 国井沙織, 森本康一, 斎藤卓也, 外村弁一郎, 赤坂一之, 日本農芸化学会大会講演要旨集,   2003 03 05
  • キウイフルーツの果肉と種子に含まれるアクチニダイン・アイソザイムの酵素反応機構の速度論的解析, 小田直子, 古田えりの, 森本康一, 井上国世, 外村ベん一郎, 日本農芸化学会大会講演要旨集,   2003 03 05
  • アテロコラーゲンのアクチニダインによる限定加水分解物の構造解析, 浜野香織, 森本康一, 斎藤卓也, 外村弁一郎, 生化学,   2002 08 25
  • 中等度好熱菌Bacillus stearothermophilus由来リシルtRNA合成酵素の結晶化とX線結晶構造解析, 森本康一, 西坂藤行, 土居賢吾, 三上文三, 滝田禎亮, 外村弁一郎, 生化学,   2002 08 25
  • Evaluation of bispecific F(ab)2 fragments prepared from mouse IgG1 monoclonal antibodies,   2002 04
  • キウイフルーツ果実の成熟過程におけるアクチニダインの果実内変動, 森本康一, 木村早苗, 水口徹, 外村弁一郎, 日本農芸化学会大会講演要旨集,   2002 03 05
  • 表面プラズモン共鳴法を用いる抗ヒスタミン抗体とヒスタミンおよびヒスタミン類縁物質との相互作用の解析, 宇野茂利, 小根田洋史, 森本康一, 藤原邦雄, 井上国世, 日本農芸化学会大会講演要旨集,   2002 03 05
  • キハダマグロ皮部由来I型コラーゲンとアクチニダイン限定分解コラーゲンの特徴, 浜野香織, 森本康一, 斎藤卓也, 外村弁一郎, 日本農芸化学会大会講演要旨集,   2002 03 05
  • キハダマグロ(Thunnus albacares)皮部の酸可溶性コラーゲン, 森本康一, 浜野香織, 外村弁一郎, 生化学,   2001 08 25
  • Optimization of immobilizing antibodies onto solid phase surfaces in sandwich enzyme immunoassay,   2001 05
  • アクチニダインによるキハダマグロ由来コラーゲンの分解, 森本康一, 仙崎暢生, 表昭宏, 外村弁一郎, 日本農芸化学会誌,   2001 03 05
  • アクチニダインの大腸菌での発現とその性質, 森本康一, 山口一生, 高橋恵, 榊利之, 井上国世, 外村弁一郎, 日本農芸化学会誌,   2001 03 05
  • アクチニダイン(Actinidia deliciosa)遺伝子の発現系の構築, 森本康一, 山口一生, 高橋恵, 榊利之, 井上国世, 外村弁一郎, 生化学,   2000 08 25
  • アクチニダイン(Actinidia deliciosa由来システインプロテアーゼ)アイソザイムの前定常状態反応の解析, 田村将也, 森本康一, 外村弁一郎, 日本農芸化学会誌,   2000 05 01
  • アクチニダイン(Actinidia deliciosa由来システインプロテアーゼ)アイソザイムの前定常状態反応の解析, 田村将也, 森本康一, 外村弁一郎, 日本農芸化学会関西支部講演会講演要旨集,   2000 02 05
  • Application of bispecific F(ab')2M fragments prepared from IgMs against carcinoembryonic antigen and alkaline phosphatase., Clinical Chemistry,   2000

Works

  • Application of tuna collagen

Research Grants & Projects

  • Application of turn collagen
  • Research on utilization of enzyme in food science
  • Research on new application of monoclonal antibody
  • Study on function and structure of fish collagen
  • Study on kinetics of enzyme reaction
  • Study on High sensitive immunoassay