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HIDAKA Yuji

    Department of Life Science　Professor
Last Updated :2024/04/25

Researcher Information

J-Global ID

Research Interests

  • フォールディング   ウログアニリン   グアニル酸シクラーゼ   結晶構造   生理活性ペプチド   ペプチドホルモン   前駆体   グアニリン   プロ領域   プロセッシング   耐熱性エンテロトキシン   ナトリウム利尿ペプチド   ペプチドホルモン前駆体   分子内シャペロン   プロペプチド   構造生物学   シャペロン   結晶化   フォールディング解析   X線結晶構造解析   フォールディング中間体   アミロイド前駆体タンパク質   プロウログアニリン   アミロイド前躯体蛋白質   

Research Areas

  • Nanotechnology/Materials / Molecular biochemistry
  • Life sciences / Structural biochemistry

Academic & Professional Experience

  • 2011 - Today  Kindai UniversityFaculty of Science and Engineering教授

Association Memberships

  • Biophysical Society (USA)   THE CHEMICAL SOCIETY OF JAPAN   The Japanese Peptide Society   

Published Papers

  • Kyona Hiroshima; Nana Sakata; Tadafumi Konogami; Shigeru Shimamoto; Yuji Hidaka
    Molecules MDPI AG 28 (23) 7754 - 7764 2023/11 [Refereed]
     
    Proopiomelanocortin (POMC) is a precursor protein of several peptide hormones, such as ACTH and β-endorphin. Almost all of the peptide hormones in POMC have been drastically investigated in terms of their biological activities. However, the biological activity of the joining peptide region (JP) in POMC is unknown. Therefore, to explore the biological activity of JP, sequence analyses of mammalian POMC were performed. We found an -Arg-Gly-Asp- (RGD) motif in several mammalian species, such as porcine, suggesting that JP has cell adhesion activity. To validate this hypothesis, the cell adhesion activities of the synthetic porcine JP peptides were examined using 293T cells. Cell adhesions were observed in a concentration-dependent manner of the JP peptides. In addition, the JP peptide competitively inhibited cell adhesion to the POMC-coated plates. Moreover, the cell adhesion activity of the joining peptide was inhibited by the addition of EDTA, indicating that the JP peptide mediates the cell adhesion activity via a receptor protein, integrin. Interestingly, a human JP peptide, which possesses an -Arg-Ser-Asp- (RSD) sequence in place of the RGD sequence, exhibited a higher ability in the cell adhesion activity than that of the porcine JP peptide, suggesting that the cell adhesion activity of the joining peptide is developed during the molecular evolution of POMC. In conclusion, our results reveal that the joining peptide in POMC plays an important role during cell adhesion and provide useful information related to signal transduction of nerve peptide hormones derived from POMC.
  • Nana Sakata; Yuri Murakami; Mitsuhiro Miyazawa; Shigeru Shimamoto; Yuji Hidaka
    Molecules 28 (8) 3494 - 3504 2023/04 [Refereed]
  • Masaya Goto; Shinya Yoshino; Kyona Hiroshima; Toru Kawakami; Kaeko Murota; Shigeru Shimamoto; Yuji Hidaka
    Molecules 28 (1) 1128 - 1142 2023/01 [Refereed]
  • Nana Sakata; Ayumi Ogata; Mai Takegawa; Yuri Murakami; Misaki Nishimura; Mitsuhiro Miyazawa; Teruki Hagiwara; Shigeru Shimamoto; Yuji Hidaka
    Molecules 27 (2) 1 - 13 2022/11 [Refereed]
  • Nana Sakata; Ayumi Ogata; Mai Takegawa; Nagisa Tajima; Misaki Nishimura; Teruki Hagiwara; Mitsuhiro Miyazawa; Shigeru Shimamoto; Yuji Hidaka
    Biochemical and Biophysical Research Communications Elsevier BV 624 35 - 39 0006-291X 2022/07 [Refereed]
  • Shigeru Shimamoto; Yuta Nakahata; Yuji Hidaka; Takuya Yoshida; Tadayasu Ohkubo
    Biomolecular NMR Assignments Springer Science and Business Media LLC 1874-2718 2022/04 [Refereed]
  • Shigeru Shimamoto; Yusuke Nakagawa; Yuji Hidaka; Takahiro Maruno; Yuji Kobayashi; Kazuki Kawahara; Takuya Yoshida; Tadayasu Ohkubo; Kosuke Aritake; Mahesh K. Kaushik; Yoshihiro Urade
    Biochemical and Biophysical Research Communications Elsevier BV 569 66 - 71 0006-291X 2021/09 [Refereed]
  • Chemical Digestion of the -Asp-Cys- Sequence for Preparation of Post-translationally Modified Proteins
    Shigeru Shimamoto; Natsumi Mitsuoka; Saki Takahashi; Tou Kawakami; Yuji Hidaka
    Protein Journal 39 (6) 711 - 716 2020/11 [Refereed][Invited]
  • Shigeru Shimamoto; Mayu Fukutsuji; Toi Osumi; Masaya Goto; Hiroshi; Toyoda; Yuji Hidaka
    Molecules 25 (20) 4798 - 4808 2020/11 [Refereed][Invited]
  • Okada S; Matsusaki M; Arai K; Hidaka Y; Inaba K; Okumura M; Muraoka T
    Chem Commun (Camb) 55 (6) 759 - 762 1359-7345 2019/01 [Refereed]
     
    Coupling of thiol and urea-type-NHC(═X)NH2 (X = O or NH) groups is effective in promoting oxidative protein folding. In particular, a thiol compound coupled with a guanidyl (X = NH) group significantly accelerates the rates of folding processes and enhances the yields of native proteins.
  • Teruki Hagiwara; Shigeru Yoshida; Yuji Hidaka
    EXPERIMENTAL LUNG RESEARCH TAYLOR & FRANCIS INC 43 (3) 150 - 157 0190-2148 2017 [Refereed]
     
    Purpose: The concentration-sensitive sodium channel (Na-C) is expressed in alveolar type II epithelial cells and pulmonary microvascular endothelial cells in mouse lungs. We recently reported that Na-C contributes to amiloride-insensitive sodium transport in mouse lungs (Respiratory Physiology & Neurobiology, 2016). However, details regarding its physiological role in the lung remain unknown. To examine whether Na-C is involved in alveolar fluid clearance during an acute lung injury (ALI), we analyzed the relationship between Na-C gene expression in the lung and the development of pulmonary edema in lipopolysaccharide (LPS)-induced ALI mice. Methods: LPS-induced ALI mice were prepared by the intratracheal administration of LPS. Bronchoalveolar lavage (BAL) neutrophils and lung water content (LWCs) were used as a marker of ALI and pulmonary edema, respectively. Na-C protein production in the lung was detected by immunoblotting and immunofluorescence. The gene expressions of Na-C and the epithelial sodium channel (ENaC) of LPS-induced ALI mice were examined by quantitative RT-PCR over a time course of 14 days. Results: The BAL neutrophil count increased until day 2 after LPS administration and had nearly recovered by day 6. LWCs in LPS-induced mice gradually increased until day 8 and had recovered by day 14. The expression of the Na-C protein in the lungs of LPS-induced mice dramatically decreased from day 2 to day 6, but recovered by day 8. The mRNA expression of Na-C decreased in the lung, as well as those for alpha-, beta-, and gamma-ENaC during ALI. Thus, Na-C expression is suppressed during the development stage of pulmonary edema and then recovers in the convalescent phase. Conclusion: Our results suggest that suppression of the gene expression of Na-C is involved in the development of pulmonary edema in ALI.
  • 清水喜久雄; 中島隆登; 松尾陽一郎; 日高雄二; 佐藤典仁; 山本幸佳
    日本放射線安全管理学会誌 日本放射線安全管理学会 15 (1) 52 - 58 1347-1503 2016/07 [Refereed]
  • Masaki Okumura; Hiroshi Kadokura; Shoko Hashimoto; Katsuhide Yutani; Shingo Kanemura; Takaaki Hikima; Yuji Hidaka; Len Ito; Kohei Shiba; Shoji Masui; Daiki Imai; Susumu Imaoka; Hiroshi Yamaguchi; Kenji Inaba
    JOURNAL OF BIOLOGICAL CHEMISTRY AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC 289 (39) 27004 - 27018 0021-9258 2014/09 [Refereed]
     
    Bisphenol A (BPA) is an endocrine disruptor that may have adverse effects on human health. We recently isolated protein-disulfide isomerase (PDI) as a BPA-binding protein from rat brain homogenates and found that BPA markedly inhibited PDI activity. To elucidate mechanisms of this inhibition, detailed structural, biophysical, and functional analyses of PDI were performed in the presence of BPA. BPA binding to PDI induced significant rearrangement of the N-terminal thioredoxin domain of PDI, resulting in more compact overall structure. This conformational change led to closure of the substrate-binding pocket in b' domain, preventing PDI from binding to unfolded proteins. The b' domain also plays an essential role in the interplay between PDI and ER oxidoreduclin 1 alpha (Ero1 alpha), a flavoenzyme responsible for reoxidation of PDI. We show that BPA inhibited Ero1 alpha-catalyzed PDI oxidation presumably by inhibiting the interaction between the b' domain of PDI and Ero1 alpha; the phenol groups of BPA probably compete with a highly conserved tryptophan residue, located in the protruding beta-hairpin of Ero1 alpha, for binding to PDI. Consistently, BPA slowed down the reoxidation of PDI and caused the reduction of PDI in HeLa cells, indicating that BPA has a great impact on the redox homeostasis of PDI within cells. However, BPA had no effect on the interaction between PDI and peroxiredoxin-4 (Prx4), another PDI family oxidase, suggesting that the interaction between Prx4 and PDI is different from that of Ero1 alpha and PDI. These results indicate that BPA, a widely distributed and potentially harmful chemical, inhibits Ero1-PDI-mediated disulfide bond formation.
  • Yuji Hidaka
    Current Protocols in Protein Science 76 (76) 28.6.1 - 6 1934-3663 2014 [Refereed][Invited]
     
    Disulfide bonds play a critical role in the maintenance of the native conformation of proteins under thermodynamic control. In general, disulfide bond formation is associated with protein folding, and this restricts the formation of folding intermediates such as misbridged disulfide isomers or kinetically trapped conformations, which provide important information related to how proteins fold into their native conformation. Therefore, numerous studies have focused on the structural analysis of folding intermediates in vitro. However, isolating or trapping folding intermediates, as well as the entire proteins, including mutant proteins, is not an easy task. Several chemical methods have recently been developed for examining peptide and protein folding and for producing, e.g., intact, post-translationally modified, or kinetically trapped proteins, or proteins with misbridged disulfide bonds. This overview introduces chemical methods for regulating the formation of disulfide bonds of peptides and proteins in the context of the thermodynamic and kinetic control of peptide and protein folding. © 2014 by John Wiley & Sons, Inc.
  • Masaki Okumura; Shigeru Shimamoto; Yuji Hidaka
    Current Protocols in Protein Science 76 (76) 28.7.1 - 13 1934-3663 2014 [Refereed][Invited]
     
    Disulfide bonds play a critical role in the folding of secretory and membrane proteins. Oxidative folding reactions of disulfide bond-containing proteins typically require several hours or days, and numerous misbridged disulfide isomers are often observed as intermediates. The rate-determining step in refolding is thought to be the disulfide-exchange reaction from nonnative to native disulfide bonds in folding intermediates, which often precipitate during the refolding process because of their hydrophobic properties. To overcome this, chemical additives or a disulfide catalyst, protein disulfide isomerase (PDI), are generally used in refolding experiments to regulate disulfide-coupled peptide and protein folding. This unit describes such methods in the context of the thermodynamic and kinetic control of peptide and protein folding, including (1) regulation of disulfide-coupled peptides and protein folding assisted by chemical additives, (2) reductive unfolding of disulfide-containing peptides and proteins, and (3) regulation of disulfide-coupled peptide and protein folding using PDI. © 2014 by John Wiley & Sons, Inc.
  • Shigeru Shimamoto; Hidekazu Katayama; Masaki Okumura; Yuji Hidaka
    Current Protocols in Protein Science 76 (76) 28.8.1 - 28 1934-3663 2014 [Refereed][Invited]
     
    Disulfide-bond formation plays an important role in the stabilization of the native conformation of peptides and proteins. In the case of multidisulfide-containing peptides and proteins, numerous folding intermediates are produced, including molecules that contain non-native and native disulfide bonds during in vitro folding. These intermediates can frequently be trapped covalently during folding and subsequently analyzed. The structural characterization of these kinetically trapped disulfide intermediates provides a clue to understanding the oxidative folding pathway. To investigate the folding of disulfide-containing peptides and proteins, in this unit, chemical methods are described for regulating regioselective disulfide formation (1) by using a combination of several types of thiol protecting groups, (2) by incorporating unique SeCys residues into a protein or peptide molecule, and (3) by combining with post-translational modification. © 2014 by John Wiley & Sons, Inc.
  • Yuji Hidaka; Shigeru Shimamoto
    Biomolecular Concepts 4 (6) 597 - 604 1868-5021 2013/12 [Refereed][Invited]
     
    Disulfide-containing proteins are ideal models for studies of protein folding as the folding intermediates can be observed, trapped, and separated by HPLC during the folding reaction. However, regulating or analyzing the structures of folding intermediates of peptides and proteins continues to be a difficult problem. Recently, the development of several techniques in peptide chemistry and biotechnology has resulted in the availability of some powerful tools for studying protein folding in the context of the structural analysis of native, mutant proteins, and folding intermediates. In this review, recent developments in the field of disulfide-coupled peptide and protein folding are discussed, from the viewpoint of chemical and biotechnological methods, such as analytical methods for the detection of disulfide pairings, chemical methods for disulfide bond formation between the defined Cys residues, and applications of diselenide bonds for the regulation of disulfide-coupled peptide and protein folding.
  • Masaki Okumura; Shigeru Shimamoto; Takeyoshi Nakanishi; Yu-ichiro Yoshida; Tadafumi Konogami; Shogo Maeda; Yuji Hidaka
    FEBS LETTERS ELSEVIER SCIENCE BV 586 (21) 3926 - 3930 0014-5793 2012/11 [Refereed]
     
    In vitro folding of disulfide-containing proteins is generally regulated by redox molecules, such as glutathione. However, the role of the cross-disulfide-linked species formed between the redox molecule and the protein as a folding intermediate in the folding mechanism is poorly understood. In the present study, we investigated the effect of the charge on a redox molecule on disulfide-coupled protein folding. Several types of aliphatic thiol compounds including glutathione were examined for the folding of disulfide-containing-proteins, such as lysozyme and prouroguanylin. The results indicate that the positive charge and its dispersion play a critical role in accelerating disulfide-coupled protein folding. (C) 2012 Federation of European Biochemical Societies. Published by Elsevier B.V. All rights reserved.
  • P. Mishra; Q. Qiu; A. Gruslin; Y. Hidaka; M. Mbikay; A. Basak
    CURRENT MOLECULAR MEDICINE BENTHAM SCIENCE PUBL LTD 12 (8) 1050 - 1067 1566-5240 2012/09 [Refereed]
     
    PC4 or PCSK4 belongs to the 9-member superfamily of mammalian subtilases collectively called Proprotein Convertases or Proprotein Convertase Subtilisin/Kexins that convert inactive precursor proteins into their active mature forms by endoproteolytic cleavage. PC4-activity plays a crucial role in mammalian fertilization via activation of sperm surface proteins. PC4 knockout mice exhibit severely impaired male fertility due to premature sperm acrosome reaction. Regulation of sperm-PC4 activity during its storage and transport through epididymis is an important determinant for ultimate egg-binding and fertilizing capacities of sperms. Herein we show that epididymal serpin CRES (cystatin related epididymal spermatogenic) recombinant protein inhibits PC4 activity in vitro in a differential manner when measured against the fluorogenic substrate Boc-RVRR-MCA depending on its oligomeric state. Thus while CRES-dimer exhibits K-i similar to 8 mu M, the corresponding monomer showed Ki > 100 mu M. Both forms also blocked PC4-mediated processing of human proIGF-2 in human placenta tropoblast cell line with dimer being more efficient. Using specific inhibitors and substrates, we also demonstrated the presence of PC4-like activity and CRES protein in varying levels in the fluids of various epididymal compartments. Our observations suggest a potential function of CRES as a regulator of PC4 in sperm-egg interaction and fertilization.
  • Yuji Hidaka
    FEBS JOURNAL WILEY-BLACKWELL 279 (13) 2261 - 2261 1742-464X 2012/07 [Refereed][Invited]
  • Masaki Okumura; Shigeru Shimamoto; Yuji Hidaka
    FEBS JOURNAL WILEY-BLACKWELL 279 (13) 2283 - 2295 1742-464X 2012/07 [Refereed][Invited]
     
    Investigations of protein folding have largely involved studies using disulfide-containing proteins, as disulfide-coupled folding of proteins permits the folding intermediates to be trapped and their conformations determined. Over the last decade, a combination of new biotechnical and chemical methodology has resulted in a remarkable acceleration in our understanding of the mechanism of disulfide-coupled protein folding. In particular, expressed protein ligation, a combination of native chemical ligation and an intein-based approach, permits specifically labeled proteins to be easily produced for studies of protein folding using biophysical methods, such as NMR spectroscopy and X-ray crystallography. A method for regio-selective formation of disulfide bonds using chemical procedures has also been established. This strategy is particularly relevant for the study of disulfide-coupled protein folding, and provides us not only with the native conformation, but also the kinetically trapped topological isomer with native disulfide bonds. Here we review recent developments and applications of biotechnical and chemical methods to investigations of disulfide-coupled peptide and protein folding. Chemical additives designed to accelerate correct protein folding and to avoid non-specific aggregation are also discussed.
  • Masatoshi Saiki; Yuji Hidaka; Masayuki Nara; Hisayuki Morii
    BIOCHEMISTRY AMER CHEMICAL SOC 51 (8) 1566 - 1576 0006-2960 2012/02 [Refereed]
     
    Prion diseases represent fatal neurodegenerative disorders caused by the aggregation of prion proteins. With regard to the formation of the amyloidogenic cross-beta-structure, the initial mechanism in the conversion to a beta-structure is critically important. To explore the core regions forming a stem of the amyloid, we designed and prepared a series of peptides comprised of two native sequences linked by a turn-inducing dipeptide moiety and examined their ability to produce amyloids. A sequence alignment of the peptides bearing the ability to form amyloid structures revealed that paired strands consisting of VNITI (residues 180-184) and VTTTT (residues 189-193) are the core regions responsible for initiating the formation of cross-beta-structures and for further ordered aggregation. In addition, most of the causative mutations responsible for inherited prion diseases were found to be located in these stem-forming regions on helix H2 and their counterpart on helix H3. Moreover, the volume effect of the nonstem domain, which contains similar to 200 residues, was deduced to be a determinant of the nature of the association such as oligomerization, because the stem-forming domain is only a small part of a prion protein. Taken together, we conclude that the mechanism underlying the initial stage of amyloidogenesis is the exposure of a newly formed intramolecular beta-sheet to a solvent through the partial transition of a native structure from an alpha-helix to a beta-structure. Our results also demonstrate that prion diseases caused by major prion proteins except the prions of some fungi such as yeast are inherent only in mammals, as evidenced by a comparison of the corresponding sequences to the stem-forming regions among different animals.
  • Masaki Okumura; Masatoshi Saiki; Hiroshi Yamaguchi; Yuji Hidaka
    FEBS JOURNAL WILEY-BLACKWELL 278 (7) 1137 - 1144 1742-464X 2011/04 [Refereed]
     
    Protein folding occurs simultaneously with disulfide bond formation. In general, the in vitro folding of proteins containing disulfide bond(s) is carried out in the presence of redox reagents, such as glutathione, to permit native disulfide pairing to occur. It is well known that the formation of a disulfide bond and the correct tertiary structure of a target protein are strongly affected by the redox reagent used. However, little is known concerning the role of each amino acid residue of the redox reagent, such as glutathione. Therefore, we prepared glutathione derivatives - glutamyl-cysteinyl-arginine (ECR) and arginyl-cysteinyl-glycine (RCG) - and examined their ability to facilitate protein folding using lysozyme and prouroguanylin as model proteins. When the reduced and oxidized forms of RCG were used, folding recovery was greater than that for a typical glutathione redox system. This was particularly true when high protein concentrations were employed, whereas folding recovery using ECR was similar to that of the glutathione redox system. Kinetic analyses of the oxidative folding of prouroguanylin revealed that the folding velocity (K(RCG) = 3.69 x 10-3 s-1) using reduced RCG/oxidized RCG was approximately threefold higher than that using reduced glutathione/oxidized glutathione. In addition, folding experiments using only the oxidized form of RCG or glutathione indicated that prouroguanylin was converted to the native conformation more efficiently in the case of RCG, compared with glutathione. The findings indicate that a positively charged redox molecule is preferred to accelerate disulfide-exchange reactions and that the RCG system is effective in mediating the formation of native disulfide bonds in proteins.
  • Yuji Hidaka; Ko-Ichi Kontani; Rina Taniguchi; Masatoshi Saiki; Sayoko Yokoi; Kenji Yukuhiro; Hiroshi Yamaguchi; Mitsuhiro Miyazawa
    BIOPOLYMERS WILEY-BLACKWELL 96 (2) 222 - 227 0006-3525 2011 [Refereed]
     
    Dragline silk is a high-performance biopolymer with exceptional mechanical properties. Artificial spider dragline silk is currently prepared by a recombinant technique or chemical synthesis. However, the recombinant process is costly and large-sized synthetic peptides are needed for fiber formation. In addition, the silk fibers that are produced are much weaker than a fiber derived from a native spider. In this study, a small peptide was chemically synthesized and examined for its ability to participate in fiber formation. A short synthetic peptide derived from Nephila clavata was prepared by a solid-phase peptide method, based on a prediction using the hydrophobic parameter of each individual amino acid residue. After purification of the spider peptide, fiber formation was examined under several conditions. Fiber formation proceeded in the acidic pH range, and larger fibers were produced when organic solvents such as trifluoroethanol and acetonitrile were used at an acidic pH. Circular dichroism measurements of the spider peptide indicate that the peptide has a 13-sheet structure and that the formation of a beta-sheet structure is required for the spider peptide to undergo fiber formation. (C) 2010 Wiley Periodicals, Inc. Biopolymers (Pept Sci) 96: 222-227, 2011.
  • Nobuyuki Fukushima; Daisuke Furuta; Yuji Hidaka; Ryutaro Moriyama; Toshifumi Tsujiuchi
    Journal of neurochemistry 109 (3) 683 - 693 0022-3042 2009 
    Many studies have shown that microtubules (MTs) interact with MT-associated proteins and motor proteins. These interactions are essential for the formation and maintenance of the polarized morphology of neurons and have been proposed to be regulated in part by highly diverse, unusual post-translational modifications (PTMs) of tubulin, including acetylation, tyrosination, detyrosination, Delta2 modification, polyglutamylation, polyglycylation, palmitoylation, and phosphorylation. However, the precise mechanisms of PTM generation and the properties of modified MTs have been poorly understood until recently. Recent PTM research has uncovered the enzymes mediating tubulin PTMs and provided new insights into the regulation of MT-based functions. The identification of tubulin deacetylase and discovery of its specific inhibitors have paved the way to understand the roles of acetylated MTs in kinesin-mediated axonal transport and neurodegenerative diseases such as Huntington's disease. Studies with tubulin tyrosine ligase (TTL)-null mice have shown that tyrosinated MTs are essential in normal brain development. The discovery of TTL-like genes encoding polyglutamylase has led to the finding that polyglutamylated MTs which accumulate during brain development are involved in synapse vesicle transport or neurite outgrowth through interactions with motor proteins or MT-associated proteins, respectively. Here we review current exciting topics that are expected to advance MT research in the nervous system.
  • Masatoshi Saiki; Mayumi Watase; Hironori Matsubayashi; Yuji Hidaka
    PROTEIN AND PEPTIDE LETTERS BENTHAM SCIENCE PUBL LTD 16 (9) 1012 - 1016 0929-8665 2009 [Refereed]
     
    Peptidylarginine deiminase IV (PAD4) catalyzes the conversion of an Arg residue to a citrulline residue in various proteins. In particular, citrullination of histone subunits, such as H2A and H3, by PAD4 is thought to be related to rheumatoid arthritis. However, the details of the citrullination mechanism of histone H2A and H3 are not yet well known. Moreover, the effects of N-terminal acetylation on histone subunits with respect to PAD4 recognition have not yet been studied. To further study the mechanism of PAD4 recognition of histone H2A and H3 subunits, a series of the N-terminal peptides was chemically synthesized and the citrullination sites were identified using MALDI-TOF/MS. N-terminal acetylation of histone H2A was not significant with respect to PAD4 recognition in vitro, but the acetylation of H3 peptide had a significant effect on PAD4 recognition in vitro, resulting in predominant citrullination at the Arg2 residue.
  • Len Ito; Yuji Hidaka; Masaki Okumura; Hironori Konishi; Hiroshi Yamaguchi
    ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY AND CRYSTALLIZATION COMMUNICATIONS BLACKWELL PUBLISHING 64 (8) 531 - 532 1744-3091 2008/06 [Refereed]
     
    Uroguanylin, which serves as an endogenous ligand of guanylyl cyclase C, is initially secreted in the form of a precursor, prouroguanylin. The N-terminal region of prouroguanylin interacts with the mature portion of prouroguanylin during the folding pathway. Here, a preliminary X-ray crystallographic study of prouroguanylin is presented. Prouroguanylin was refolded, purified and crystallized using the hanging-drop vapour-diffusion method. Prouroguanylin crystals were cryocooled and used for data collection. The diffraction data showed that the crystals belonged to space group P6(1)22, with unit-cell parameters a = b = 55.6, c = 157.7 angstrom, and diffracted to 2.5 angstrom resolution. The structure is currently being analyzed.
  • Len Ito; Yuji Hidaka; Masaki Okumura; Hironori Konishi; Knut Adermann; Hiroshi Yamaguchi
    Acta Crystallographica Section F: Structural Biology and Crystallization Communications BLACKWELL PUBLISHING 64 (8) 771 - 771 1744-3091 2008 [Refereed]
  • Tsuyoshi Imasaki; Toshiyuki Shimizu; Hiroshi Hashimoto; Yuji Hidaka; Shingo Kose; Naoko Imamoto; Michiyuki Yamada; Mamoru Sato
    MOLECULAR CELL CELL PRESS 28 (1) 57 - 67 1097-2765 2007/10 [Refereed]
     
    Transportin 1 (Trn1) is a transport receptor that transports substrates from the cytoplasm to the nucleus through nuclear pore complexes by recognizing nuclear localization signals (NLSs). Here we describe four crystal structures of human Trn1 in a substrate-free form as well as in the complex with three NLSs (hnRNP D, JKTBP, and TAP, respectively). Our data have revealed that (1) Trn1 has two sites for binding NLSs, one with high affinity (site A) and one with low affinity (site B), and NLS interaction at site B controls overall binding affinity for Trn1; (2) Trn1 recognizes the NLSs at site A followed by conformational change at site B to interact with the NLSs; and (3) a long flexible loop, characteristic of Trn1, interacts with site 13, thereby displacing transport substrate in the nucleus. These studies provide deep understanding of substrate recognition and dissociation by Trn1 in import pathways.
  • Imasaki T; Shimizu T; Hashimoto H; Hidaka Y; Yamada M; Sato M
    Acta Crystallogr. Sect. F 62 (Pt 8) 785 - 787 2006/08 [Refereed]
     
    Nucleocytoplasmic transport of proteins with molar masses of larger than 60 000 is mediated by transport receptors. The transport receptor transportin1 (Trn1) transports various kinds of RNA-binding proteins such as JKTBP, hnRNP D and TAP. Trn1 was successfully cocrystallized with nucleocytoplasmic shuttling fragments of JKTBP and hnRNP D and a nuclear localization fragment of TAP. The crystal of the Trn1–JKTBP fragment complex belongs to space group P21212, with unit-cell parameters a = 131.5, b =171.5, c = 68.2 Å. The crystals of Trn1 in complex with hnRNP D and TAP fragments are orthorhombic, space group P212121, with unit-cell parameters a = 69.1, b = 119.1, c = 151.1 Å and a = 69.0, b = 119.1, c = 146.0 Å, respectively. The crystals diffracted to beyond 3.0, 3.2 and 2.4 Å resolution, respectively, using synchrotron radiation at SPring-­8.
  • K Arita; T Shimizu; H Hashimoto; Y Hidaka; M Yamada; M Sato
    PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA NATL ACAD SCIENCES 103 (14) 5291 - 5296 0027-8424 2006/04 [Refereed]
     
    Histone arginine methylation is a posttranslational modification linked to the regulation of gene transcription. Unlike other posttranslational modifications, methylation has generally been regarded as stable, and enzymes that demethylate histone arginine residues have not been identified. However, it has recently been shown that human peptidylarginine deiminase 4 (PAD4), a Ca2+-dependent enzyme previously known to convert arginine residues to citrulline in histones, can also convert monomethylated arginine residues to citrulline both in vivo and in vitro. Citrullination of histone arginine residues by the enzyme antagonizes methylation by histone arginine methyltransferases and is thus a novel posttranslational modification that regulates the level of histone arginine methylation and gene activity. Here we present the crystal structures of a Ca2+-bound PAD4 mutant in complex with three histone N-terminal peptides, each consisting of 10 amino acid residues that include one target arginine residue for the enzyme (H3/Arg-8, H3/Arg-17, and H4/Arg-3). To each histone N-terminal peptide, the enzyme induces a beta-turn-like bent conformation composed of five successive residues at the molecular surface near the active site cleft. The remaining five residues are highly disordered. The enzyme recognizes each peptide through backbone atoms of the peptide with a possible consensus recognition motif. The sequence specificity of the peptide recognized by this enzyme is thought to be fairly broad. These observations provide structural insights into target protein recognition by histone modification enzymes and illustrate how PAD4 can target multiple arginine sites in the histone N-terminal tails.
  • Makoto Hasegawa; Yoshiko Matsumoto-Ishikawa; Atsushi Hijikata; Yuji Hidaka; Mitiko Go; Yasutsugu Shimonishi
    Protein Journal 24 (5) 315 - 325 1572-3887 2005/07 [Refereed]
     
    Guanylyl cyclase C (GC-C) is a single-transmembrane receptor that is specifically activated by endogenous ligands, including guanylin, and the exogenous ligand, heat-stable enterotoxin. Using combined HPLC separation and MS analysis techniques the positions of the disulfide linkages in the extracellular ligand-binding domain (ECD) of GC-C were determined to be between Cys7-Cys94, Cys72-Cys77, Cys 101-Cys128 and Cys179-Cys226. Furthermore, a three-dimensional structural model of the ECD was constructed by homology modeling, using the structure of the ECD of GC-A as a template (van den Akker et al., 2000, Nature, 406: 101-104) and the information of the disulfide linkages. Although the GC-C model was similar to the known structure of GC-A, importantly its ligand-binding site appears to be located on the quite different region from that in GC-A. © 2005 Springer Science+Business Media, Inc.
  • A Schulz; UC Marx; N Tidten; T Lauber; Y Hidaka; K Adermann
    JOURNAL OF PEPTIDE SCIENCE WILEY-BLACKWELL 11 (6) 319 - 330 1075-2617 2005/06 [Refereed]
     
    The peptide hormones guanylin and uroguanylin are ligands of the intestinal guanylyl cyclase-C (GC-C) that is involved in the regulation of epithelial water and electrolyte transport. The small peptides contain 15 and 16 amino acids, respectively, and two disulfide bonds with a 1-3/2-4 connectivity. This structural feature causes the unique existence of two topological isoforms for each peptide in an approximate 3: 2 ratio, with only one of the isoforms exhibiting GC-C-activating potential. The two uroguanylin isomers can be separated by HPLC and are of sufficient stability to be studied separately at ambient temperatures while the two guanylin isomers are rapidly interconverting even at low temperatures. Both isomers show clearly distinguishable (1)H chemical shifts. To investigate the influence of certain amino acid side chains on this isomerism and interconversion kinetics, derivatives of guanylin and uroguanylin (L-alanine scan and chimeric peptides) were designed and synthesized by Fmoc solid-phase chemistry and compared by HPLC and 2D (1)H NMR spectroscopy. Amino acid residues with the most significant effects on the interconversion kinetics were predominantly identified in the COOH-terminal part of both peptides, whereas amino acids in the central part of the peptides only moderately affected the interconversion. Thus, the conformational conversion among the isomers of both peptides is under the control of a COOH-terminal sterical hindrance, providing a detailed model for this dynamic isomerism. Our results demonstrate that kinetic control of the interconversion process can be achieved by the introduction of side chains with a defined sterical profile at suitable sequence positions. This is of potential impact for the future development of GC-C peptide agonists and antagonists. Copyright (c) 2004 European Peptide Society and John Wiley & Sons, Ltd.
  • T Hagiwara; Y Hidaka; M Yamada
    BIOCHEMISTRY AMER CHEMICAL SOC 44 (15) 5827 - 5834 0006-2960 2005/04 [Refereed]
     
    Interplay of various covalent modifications of histone tails has an essential role in regulation of chromatin function. Peptidylarginine deiminase (PADI) 4 deiminates protein arginine to citrulline in a Ca2+-dependent manner and is present in the nucleus of granulocyte-differentiated HL-60 cells. When these cells are treated with the calcium ionophore A23187, core histone deimination occurs. To determine the deimination sites of histones, histone species were purified by reverse-phase high-performance liquid chromatography (RP-HPLC) from the cells. Immunoblotting using antimodified citrulline antibody indicated that histones H2A, H3, and H4 but not H2B were deiminated. H2A and H4 were digested with Staphylococcus aureus V8 protease, and the digests were separated by RP-HPLC. Immuno dot-blotting and mass spectrometry showed that the deiminated residues were present in H2A (1-56) and H4 (1-52) regions but not in other regions. The H2A peptide (1-56) was digested with alpha-chymotrypsin, and the deiminated peptide was separated from the corresponding nondeiminated peptide by RP-HPLC. The deiminated residue was found to be limited to residues 1-23. Similarly, digestion of the H4 peptide (1-52) with endoproteinase Asp-N and separation of the deiminated peptide from the nondeiminated peptide indicated that the deiminated residue was limited to residues 1-23. Mass spectrometry of lysylendopeptidase digests of the H2A (1-23) and H4 (1-23) peptides showed that deimination occurred at arginine 3 of the N-terminal sequence Ac-SGRGK common to H2A and H4. These results suggest that PADI4 deiminates only a restricted site of target proteins in cells. Deimination of histones is discussed in relation to chromatin structure and function.
  • Side Chain Contributions to the Interconversion of the Topological Isomers of Guanylin-like Peptides
    Axel Schulz; Ute Marx; Naomi Tidten; Thomas Lauber; Yuji Hidaka; Knut Adermann
    Journal of Peptide Science 11 319 - 330 2004/11 [Refereed]
  • The Micro Domain Responsible for Ligand-binding of Guanylyl Cyclase C
    Yuji Hidaka; Yoshiko Matsumoto; Yasutsugu Shimonishi
    FEBS Lett 526 58 - 62 2002/07 [Refereed]
  • Y Hidaka; C Shimono; M Ohno; N Okumura; K Adermann; WG Forssmann; Y Shimonishi
    JOURNAL OF BIOLOGICAL CHEMISTRY AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC 275 (33) 25155 - 25162 0021-9258 2000/08 [Refereed]
     
    Guanylyl cyclase activating peptide II (GCAP-II), an endogenous ligand of guanylyl cyclase C, is produced via the processing of the precursor protein (prepro-GCAP-II), We have previously shown that the propeptide in pro-GCAP-II functions as an intramolecular chaperone in the proper folding of the mature peptide, GCAP-II (Hidaka, Y., Ohno, M., Hemmasi, B,, Hill, O., Forssmann, W.-G., and Shimonishi, Y. (1998) Biochemistry 37, 8498-8507). Here, we report an essential region in pro-GCAP-II for the correct disulfide pairing of the mature peptide, GCAP-II. Five mutant proteins, in which amino acid residues were sequentially deleted from the N terminus, and three mutant proteins of pro-GCAP-II, in which N-terminal 6, 11, or 17 amino acid residues were deleted, were overproduced using Escherichia coli or human kidney 293T cells respectively, Detailed analysis of in vivo or in vitro folding of these mutant proteins revealed that one or two amino acid residues at the N terminus of pro-GCAP-II are critical, not only for the chaperone function in the folding but also for the net stabilization of pro-GCAP-II. In addition, size exclusion chromatography revealed that pro-GCAP-II exists as a dimer in solution. These data indicate that the propeptide has two roles in proper folding: the disulfide-coupled folding of the mature region and the dimerization of pro-GCAP-II.
  • M Hasegawa; Y Hidaka; Y Matsumoto; T Sanni; Y Shimonishi
    JOURNAL OF BIOLOGICAL CHEMISTRY AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC 274 (44) 31713 - 31718 0021-9258 1999/10 [Refereed]
     
    Guanylyl cyclase C, one of the family of membrane-bound guanylyl cyclases, consists of an extracellular domain and an intracellular domain, which are connected by a single transmembrane polypeptide. The extracellular domain binds unique small polypeptides with high specificity, which include the endogenous peptide hormones, guanylin and uroguanylin, as well as an exogenous enterotoxigenic peptide, heat-stable enterotoxin, secreted by pathogenic Escherichia coli. Information on this specific binding is propagated into the intracellular domain, followed by the synthesis of cGMP, a second messenger that regulates a variety of intracellular physiological processes. This study reports the design of a photoaffinity labeled analog of heat-stable enterotoxin (biotinyl-(AC(5))(2)-[Gly(4),Pap(11)]STp(4-17)), which incorporates a Pap residue (p-azidophenylalanine) at position 11 and a biotin moiety at the N terminus, and the use of this analog to determine the ligand-binding region of the extracellular domain of guanylyl cyclase C, The endoproteinase Lys-C digestion of the extracellular domain, which was covalently labeled by this ligand, and mass spectrometric analyses of the digest revealed that the ligand specifically binds to the region (residue 387 to residue 393) of guanylyl cyclase C. This region is localized close to the transmembrane portion of guanylyl cyclase C on the external cellular surface. This result was further confirmed by characterization of site-directed mutants of guanylyl cyclase C in which each amino acid residue was substituted by an Ala residue instead of residues normally located in the region. This experiment provides the first direct demonstration of the ligand-binding site of guanylyl cyclase C and will contribute toward an understanding of the receptor recognition of a ligand and the modeling of the interaction of the receptor and its ligand at the molecular level.
  • M Hasegawa; Y Hidaka; A Wada; T Hirayama; Y Shimonishi
    EUROPEAN JOURNAL OF BIOCHEMISTRY BLACKWELL SCIENCE LTD 263 (2) 338 - 345 0014-2956 1999/07 [Refereed]
     
    The role of carbohydrate moieties at the N-linked glycosylation sites of guanylate cyclase C (GC-C), a receptor protein for guanylin, uroguanylin and heat-stable enterotoxin, in ligand binding and structural stability was examined using site-directed mutagenesis of the putative N-linked glycosylation sites in the extracellular domain (ECD) of porcine GC-C. For this purpose, eight mutant proteins of ECD (N9A, N20A, N56A, N172A, N261A, N284A, N334A and N379A): and six mutant proteins of the complete GC-C (N9A, S11A, N172A, T174A, N379A and T381A) were prepared, in which Ala replaced Asn, Ser and Thr at the N-linked glycosylation consensus sites. All the mutant proteins showed a ligand-binding affinity (K-d) similar to those of the wild-type proteins, although the deletion of a carbohydrate moiety at each of the N-linked glycosylation sites affected the ligand-binding ability of ECD or GC-C to some degree. However, the mutant proteins of ECD (N379A) and GC-C (N379A and T381A) showed considerably decreased binding ability in the context of maximum capacity (B-max to a ligand, despite the fact that the expression levels of these mutant proteins were nearly the same as the wild-type proteins. Moreover, the mutant protein of ECD (N379A) was considerably less stable to a denaturant. These results clearly indicate a crucial role for the carbohydrate moiety at N379, which is located near the transmembrane region, in structural stability, the ability to bind to a ligand and the cyclase catalytic activity of GC-C, and provide a route for the elucidation of the mechanism of the interaction between GC-C and a ligand.
  • Axel Schulz; Ute C. Marx; Yuji Hidaka; Yasutsugu Shimonishi; Paul Rösch; Wolf-Georg Forssmann; Knut Adermann
    Protein Science Cambridge University Press 8 (9) 1850 - 1859 0961-8368 1999 [Refereed]
     
    Guanylin is a guanylyl cyclase (GC)-activating peptide that is mainly secreted as the corresponding prohormone of 94 amino acid residues. In this study, we show that the originally isolated 15-residue guanylin, representing the COOH-terminal part of the prohormone, is released from the prohormone by cleavage of an Asp-Pro amide bond under conditions applied during the isolation procedures. Thus, the 15-residue guanylin is probably a non-native, chemically induced GC-activating peptide. This guanylin molecule contains two disulfide bonds that are absolutely necessary for receptor activation. We demonstrate that the folding of the reduced 15-residue guanylin results almost completely in the formation of the two inactive disulfide isomers. In contrast, the reduced form of proguanylin containing the entire prosequence folds to a product with the native cysteine connectivity. Because proguanylin lacking the 31 NH2-terminal residues of the prosequence folds only to a minor extent to guanylin with the native disulfide bonds, it is evident that this NH2-terminal region contributes significantly to the correct disulfide- coupled folding. Structural studies using CD and NMR spectroscopy show that native proguanylin contains a considerable amount of α-helical and, to a lesser extent, β-sheet structural elements. In addition, a close proximity of the NH2- and the COOH-terminal regions was found by NOESY. It appears that this interaction is important for the constitution of the correct conformation and provides an explanation of the minor guanylyl cyclase activity of proguanylin by shielding the bioactive COOH-terminal domain from the receptor.
  • T Akao; S Hashimoto; K Kobashi; Y Hidaka
    JOURNAL OF BIOCHEMISTRY JAPANESE BIOCHEMICAL SOC 125 (1) 27 - 30 0021-924X 1999/01 [Refereed]
     
    A peptide has been isolated from pronase digest of bovine serum albumin as the stimulatory factor of streptolysin S (SLS) production by Streptococcus pyogenes, and its primary structure has been deduced [Akao et at. (1992) Infect. Immun. 60, 4777-4780], To determine the essential structure for the stimulation, a peptide (P-l) having the deduced structure, in which three peptide fragments are linked by two disulfide bonds, and shorter analogs (P-2 to P-4) of peptide P-l were chemically synthesized. Another peptide (P-5), in which Ala is inserted between the two Cys residues in the middle peptide chain of P-1, was also synthesized, These synthetic peptides were identified by mass spectrometry and analysis of amino acid compositions, The synthetic P-1 stimulated SLS production in a dose-dependent manner. Other peptide analogs also showed remarkable stimulation of SLS production. Treatment of P-l with performic acid resulted in loss of its stimulatory activity, indicating that disulfide bridges of the peptides are necessary for their activity on SLS production, These results suggest that the unique primary structure of three peptide chains linked by two disulfide bridges is requisite for the stimulatory effect on SLS production.
  • Makoto Hasegawa; Yuki Kawano; Yoshiko Matsumoto; Yuji Hidaka; Jun-Ichi Fujii; Naoyuki Taniguchi; Akihiro Wada; Toshiya Hirayama; Yasutsugu Shimonishi
    Protein Expression and Purification Academic Press Inc. 15 (3) 271 - 281 1046-5928 1999 [Refereed]
     
    Guanylyl cyclase (GC)-C, a single-transmembrane receptor protein for heat-stable enterotoxin, guanylin, and uroguanylin, and its N-terminal extracellular domain were prepared at a high level of expression from a system constructed of Sf21 insect cells and recombinant baculovirus. The recombinant GC-C, containing the complete sequence, retained its binding affinity to heat-stable enterotoxin with a K(D) value (6.2 x 10-10 M) and cyclase catalytic activity at a level similar to those of GC-C expressed in mammalian cell lines, such as COS-7. The N-terminal extracellular domain was prepared in a form which contained the hexahistidine tail at its C-terminus and was purified as a homogenous protein by Con A and Ni-chelating affinity chromatography from the culture medium of the insect cells. The purified N-terminal extracellular domain of GC-C exhibited the high (K(D) = 4 x 10-10 M) and low (K(D) = 7 x 10-8 M) affinity sites in binding to heat-stable enterotoxin. These results clearly indicate that the N-terminal extracellular domain of GC-C possesses the same biochemical characteristics as the complete GC-C protein even in the membrane-free form. Moreover, the extracellular domain is able to form an oligomer in a ligand-dependent manner, suggesting that the N-terminal extracellular domains interact with one another in binding to ligands.
  • In vitro disulfide-coupled folding of guanylyl cyclase-activating peptide and its precursor protein.
    Yuji Hidaka; Megumu Ohno; Bahram Hemmasi; Oliver Hill; Wolf-Georg Forssmann; Yasutsugu Shimonishi
    Biochemistry 37 8498 - 8507 1998/05 [Refereed]
  • K Oishi; B Sar; A Wada; Y Hidaka; S Matsumoto; H Amano; F Sonoda; S Kobayashi; T Hirayama; T Nagatake; K Matsushima
    INFECTION AND IMMUNITY AMER SOC MICROBIOLOGY 65 (7) 2648 - 2655 0019-9567 1997/07 [Refereed]
     
    Persistent infection with Pseudomonas aeruginosa increases interleukin-8 (IL-8) levels and causes dense neutrophil infiltrations in the airway of patients with chronic airway diseases. To investigate the role of. aeruginosa infection in IL-8 production in the airway of these patients, eve examined whether cell lysates of P. aeruginosa could cause IL-S production from human bronchial epithelial cells. Diluted sonicated supernatants of P. aeruginosa (SSPA) vith a mucoid or nonmucoid phenotype stimulated human bronchial epithelial (BET-IA) cells to produce IL-8. In this study, we have purified a 59-kDa heat-stable protein with IL-8-inducing activity from the SSPA by sequential ion-exchange chromatography. The N-terminal sequence of this purified protein completely matched a sequence at the N-terminal part of the mature protein of nitrite reductase from P. aeruginosa. In addition, immunoblotting with a polyclonal immunoglobulin G (IgG) against recombinant Pseudomonas nitrile reductase demonstrated a specific binding to the purified protein. Furthermore, the immunoprecipitates of the SSPA with a polyclonal IgG against recombinant nitrite reductase induced a twofold-higher IL-8 production in the BHT-IA cell culture than did the immunoprecipitates of the SSPA with a control IgG. These lines of evidence confirmed that Pseudomonas nitrite reductase was responsible for IL-8 production in the BET-1A cells. Tie purified nitrite reductase induced maximal expression of IL-8 mRNA in the BET-1A cells at 1 to 3 h after stimulation, and the IL-IS mRNA expression declined by 8 h after stimulation. New protein translation was not required far nitrite reductase-mediated IL-S mRNA expression in the BET-1A cells. Nitrite reductase stimulated the BET-1A cells, as well as human alveolar macrophages, pulmonary fibroblasts, and neutrophils, to produce IL-8. In contrast, nitrite reductase induced significant levels of tumor necrosis factor alpha and IL-1 beta protein only in human alveolar macrophages. These data support the notion that nitrite reductase from P. aeruginosa induces the production of inflammatory cytokines by respiratory cells and may contribute to the pathogenesis of chronic airway diseases and persistent P. aeruginosa infection.
  • Makoto Hasegawa; Yuki Kawano; Kazuya Matsumoto; Yuji Hidaka; Takashi Sato; Yasutsugu Shimonishi
    Letters in Peptide Science Springer Science and Business Media LLC 4 (1) 1 - 11 0929-5666 1997/03
  • Demonstration of dimer formation of the cytoplasmic domain of a transmembrane osmosensor protein, EnvZ, of escherichia coli using Ni-histidine tag affinity chromatography
    Yuji Hidaka; Heiyoung Park; Masayori Inouye
    FEBS Lett 400 238 - 242 1997/02 [Refereed]
  • Makoto Hasegawa; Yuki Kawano; Kazuya Matsumoto; Yuji Hidaka; Takashi Sato; Yasutsugu Shimonishi
    International Journal of Peptide Research and Therapeutics Springer New York 4 (1) 1 - 11 1573-3904 1997 [Refereed]
     
    The heat-stable enterotoxin (ST), produced by Escherichia coli, causes acute diarrhea in infants and domestic animals by activation of the intestinal membrane-bound receptor, guanylyl cyclase C. We have investigated a region on the ST molecule, which is recognized by the receptor, by introducing a photochromophore, p-azidophenylalanine (Pap), into three different regions of STp(4-17), which has the full toxic activity. Each ST analog bound to the receptor, but only STp(4-17) containing a Pap residue at position 11 in the central portion of the ST molecule, showed a high efficiency in the reaction which cross-linked with the receptor by UV radiation. These data clearly demonstrate that the region of the ST molecule encompassing the Asn11 residue directly interacts with the receptor. © 1997 ESCOM Science Publishers B.V.
  • A Wada; T Hirayama; H Kitaura; JI Fujisawa; M Hasegawa; Y Hidaka; Y Shimonishi
    INFECTION AND IMMUNITY AMER SOC MICROBIOLOGY 64 (12) 5144 - 5150 0019-9567 1996/12 [Refereed]
     
    Guanylyl cyclase C (STaR), a receptor protein for heat-stable enterotoxin (STa) elaborated by Escherichia coli, is associated with and spans the plasma membrane of mammalian intestinal cells. The extracellular domain functions in the binding of STa and the association of each domain to an oligomeric form. Two amino acid residues, Arg-136 and Asp-347, were identified as the residues binding to STa in the extracellular domain of pig STaR by site-directed mutagenesis and analysis of expression on 293T cells. Replacement of these residues bg other amino acid residues resulted in the toss of binding of pig STaR to STa, and as a result, STa-induced guanylyl cyclase activity was eliminated. Furthermore, mutation in a region (from Asp-347 to Val-401) which is close to the transmembrane domain caused a significant reduction in both STa-binding activity and guanylyl cyclase catalytic activity. These results suggest that the region adjacent to the transmembrane domain plays an important role in facilitating a favorable conformation of STaR for STa binding.
  • Akihiro Wada; Makoto Hasegawa; Kazuya Matsumoto; Takuro Niidome; Yuki Kawano; Yuji Hidaka; Philip Ian Padilla; Hisao Kurazono; Yasutsugu Shimonishi; Toshiya Hirayama
    FEBS Letters Elsevier 384 (1) 75 - 77 0014-5793 1996/04 [Refereed]
     
    To characterize Ser1029 in STaR at a consensus sequence of phosphorylation site by PKC, two mutants of mS1029A with replacement of Ser1029 to Ala1029 and CΔ1029 lacking 22 amino acids including Ser1029 were prepared. Preincubation of the wild type-STaR (wt-STaR) transfectant with 1 μM PMA caused additional STa-mediated guanylyl cyclase (GC) activation compared to control, whereas the mS1029A- and CΔ1029-transfected cells did not show a similar enhanced GC activation by PMA. After metabolic labeling with [32P]phosphate, transfected cells with wt-STaR and mutants were incubated with 1 μM PMA. Subsequent 32P-radiolabeled proteins were immunoprecipitated using anti-STaR antibody, and analyzed by autoradiography after separation on SDS-PAGE. The immunoprecipitated wt-STaR but not mS1029A and CΔ1029 had a significant radioactivity. These results suggest that the effect of PMA on wt-STaR transfectants may be caused by phosphorylation of Ser1029. The CΔ1012 mutant, with further truncation (Gln1012-Phe1050) of the carboxy terminus, did not show STa-mediated GC activation. Based on these data, these 17 amino acids (Gln1012-Ala1028), essential for signaling of GC activation by STa, have an abundance of basic amino acids which might be functionally influenced by phosphorylation of Ser1029.
  • Masahiko Ehara; Yoshio Ichinose; Toshiya Hirayama; Hisao Kurazano; Yuji Hidaka; Kouichi Morita; Akira Igarashi; Shouichi Shimodori
    FEMS Microbiology Letters Oxford University Press (OUP) 123 (1-2) 185 - 191 1574-6968 1994/10 [Refereed]
     
    The gene encoding an 18 kDa fimbrial subunit of Vibrio cholerae O1 was identified in a fimbriate strain Bgd17. Mixed oligoprimers were prepared based on the amino acid sequence of the N-terminus and that from a cyanogen bromide-cleaved fragment of the fimbrillin. A PCR-amplified 185 bp DNA fragment was sequenced. This 185 bp fragment was further extended to 540 bp to 3′ and 5′ termini by RNA-PCR using a primer containing a random hexamer at its 3′ end. This fragment did not contain the stop codons. It was further extended by a gene walking method using EcoRI cassette and its primers. Finally a 660 bp fragment was obtained and sequenced. This fragment contained the complete open reading frame of the structural subunit of the fimbriae, composed of 169 amino acids with a molecular mass of 17435.65 and a leader sequence of 6 or 9 amino acids. The deduced amino acid sequence of the polypeptide encoded by the gene, designated fimA, displayed a highly conserved sequence of MKXXXGFTLI EL of type 4 fimbriae. © 1994.
  • Significant amino-acids of Heat-stable Enterotoxin Receptor (StaR) for STa-binding
    Akihiro Wada, A; Oshita, T; Kitaura, H; Jun-ichi Fujisawa; Yuji Hidaka; Yasutsugu Shimonishi; Toshiya Hirayama
    Japanese Journal of Medical Science & Biology 47 315 - 316 1994/04 [Refereed]
  • T AKI; K ONO; Y HIDAKA; Y SHIMONISHI; T JYO; T WADA; M YAMASHITA; S SHIGETA; Y MUROOKA; S OKA
    INTERNATIONAL ARCHIVES OF ALLERGY AND IMMUNOLOGY KARGER 103 (4) 357 - 364 1018-2438 1994/04 [Refereed]
     
    Two IgE epitope sequences comprising Ser(56)-Pro-Val-Thr-Lys-Arg-Ala-Ser-Leu-Lys-Ile-Asp-Ser-Lys-Lys(70) and Asp(104)-Val-Glu-Leu-Ser-Leu-Arg-Ser-Ser-Asp-Ile-Ala(115) were identified by deletion analysis of the cDNA encoding a new 39-kD protein of mite allergen. A synthetic dodecapeptide corresponding to the latter epitope sequence functioned as a monovalent and mite-specific hapten. Replacement of each of the 12 amino acid residues with Gly, using site-directed mutagenesis, indicated that Arg(110) may play a central role in IgE binding. However, the 8 allergic sera tested exhibited a wide variation in their amino acid residues required for reactivity to IgE in allergic sera.
  • A WADA; T HIRAYAMA; S KITAO; J FUJISAWA; Y HIDAKA; Y SHIMONISHI
    MICROBIOLOGY AND IMMUNOLOGY CENTER ACADEMIC PUBL JAPAN 38 (7) 535 - 541 0385-5600 1994 [Refereed]
     
    A cDNA encoding the receptor protein for a heat-stable enterotoxin (STa) produced by enterotoxigenic Eschrerichia coli was cloned from intestinal epithelial cells of a 10-week-old pig. The cDNA had an open reading frame of 3,219 base pairs and coded for a protein with 1,073 amino acid residues. The mature protein consisted of 1,050 amino acid residues with a molecular mass of ca. 121 kDa and was 87% and 82% identical with the human and rat protein, respectively. The CHO cell line overexpressing the pig recombinant STa receptor specifically bound to a photoaffinity-labeled analog of STa and showed marked elevation of the cellular content of cGMP in response to STa.
  • High-Performance Liquid Chromatographic Determination of Neutral and Amino Monosaccharides by Ultraviolet and Fluorescence Detection of Sugar 9-Fluorenylmethoxycarbonyl Hydrazones and 9-Fluorenylmethoxycarbonyl Amino Sugars at Picomole and Sub-Picomole Le
    Renen Zhang; Zhidan Zhang; Guoquan Liu; Yuji Hidaka; Yasutsugu Shimonishi
    Journal of Chromatography, 646 45 - 52 1993/03 [Refereed]
  • Toshiya Hirayama; Akihiro Wada; Yuji Hidaka; Jun-Ichi Fujisawa; Yoshifumi Takeda; Yasutsugu Shimonishi
    Microbial Pathogenesis 15 (4) 283 - 291 1096-1208 1993 [Refereed]
     
    A fragment of guanylate cyclase C (GC-C) of about 1.7 k bp corresponding to amino acids 1-553 spanning the extracellular and transmembrane domains and a portion of the intracellular region was amplified using template cDNA prepared from rat intestinal cells by the polymerase chain reaction method. The cloned 1.7 k bp fragment was inserted into the mammalian expression vector pCGUT and the truncated GC-C expressed on the surface of COS-7 cells was demonstrated to bind heat-stable enterotoxin by photo affinity labeling with 125 I-N-5-azidonitrobenzoyl-STh[5-19]. Analysis by sodium dodecyl sulfate-polyacrylamide disc gel electrophoresis showed that the truncated GC-C formed dimers on the surface of COS-7 cells. The intracellular region of GC-C was found not to be necessary for dimer formation by the GC-C. Comparison of the molecular weights of the truncated GC-C expressed in COS-7 cells and Escherichia coli suggested that the truncated GC-C was glycosylated in the mammalian expression system. © 1993 Academic Press. All rights reserved.
  • Y HIDAKA; K OHMORI; A WADA; H OZAKI; H ITO; T HIRAYAMA; Y TAKEDA; Y SHIMONISHI
    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS 176 (3) 958 - 965 0006-291X 1991/05 [Refereed]
  • H OZAKI; H KUBOTA; T SATO; Y HIDAKA; H TAMAOKI; Y KOBAYASHI; Y KYOGOKI; T SUGIMURA; A TAI; Y SHIMONISHI
    BULLETIN OF THE CHEMICAL SOCIETY OF JAPAN CHEMICAL SOC JAPAN 64 (4) 1136 - 1144 0009-2673 1991 [Refereed]
     
    Heat-stable enterotoxin (ST(p)) produced by a porcine strain of enterotoxigenic Escherichia coli is a peptide of 18 amino acid residues. The full toxicity is generated by a core peptide of 13 amino acids from residue 5 to 17 (ST(p)(5-17)). Detailed conformational analysis of ST(p)(5-17) was performed by NMR spectroscopy and distance geometry calculations. The tetriary structure of ST(p)(5-17) was found to have a right-handed spiral fold throughout the whole molecule which was stabilized by the network of disulfide linkages and hydrogen bonds. This folding pattern was demonstrated in both organic and aqueous solutions.
  • S YAMASAKI; T SATO; Y HIDAKA; H OZAKI; H ITO; T HIRAYAMA; Y TAKEDA; T SUGIMURA; A TAI; Y SHIMONISHI
    BULLETIN OF THE CHEMICAL SOCIETY OF JAPAN CHEMICAL SOC JAPAN 63 (7) 2063 - 2070 0009-2673 1990/07 [Refereed]
  • Y HIDAKA; K SATO; H NAKAMURA; J KOBAYASHI; Y OHIZUMI; Y SHIMONISHI
    FEBS LETTERS ELSEVIER SCIENCE BV 264 (1) 29 - 32 0014-5793 1990/05 [Refereed]
  • Y HIDAKA; Y SHIMONISHI
    BULLETIN OF THE CHEMICAL SOCIETY OF JAPAN CHEMICAL SOC JAPAN 62 (6) 1986 - 1994 0009-2673 1989/06 [Refereed]
  • H KUBOTA; Y HIDAKA; H OZAKI; H ITO; T HIRAYAMA; Y TAKEDA; Y SHIMONISHI
    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS ACADEMIC PRESS INC JNL-COMP SUBSCRIPTIONS 161 (1) 229 - 235 0006-291X 1989/05 [Refereed]
  • S YAMASAKI; Y HIDAKA; H ITO; Y TAKEDA; Y SHIMONISHI
    BULLETIN OF THE CHEMICAL SOCIETY OF JAPAN CHEMICAL SOC JAPAN 61 (5) 1701 - 1706 0009-2673 1988/05 [Refereed]
  • Y HIDAKA; H KUBOTA; S YOSHIMURA; H ITO; Y TAKEDA; Y SHIMONISHI
    BULLETIN OF THE CHEMICAL SOCIETY OF JAPAN CHEMICAL SOC JAPAN 61 (4) 1265 - 1271 0009-2673 1988/04 [Refereed]
  • S YOSHIMURA; Y HIDAKA; S AIMOTO; Y SHIMONISHI; T TAKEDA; T MIWATANI; Y TAKEDA
    BULLETIN OF THE CHEMICAL SOCIETY OF JAPAN CHEMICAL SOC JAPAN 60 (7) 2481 - 2489 0009-2673 1987/07 [Refereed]
  • Y SHIMONISHI; Y HIDAKA; M KOIZUMI; M HANE; S AIMOTO; T TAKEDA; T MIWATANI; Y TAKEDA
    FEBS LETTERS ELSEVIER SCIENCE BV 215 (1) 165 - 170 0014-5793 1987/05 [Refereed]

Conference Activities & Talks

  • Understanding the mechanism by which PDI family catalyze oxidative folding of precursor protein  [Not invited]
    増田光紀; 木下 岬; 山口 宏; 日高雄二; 稲葉謙次; 奥村正樹
    第92回日本生化学会大会  2019/09
  • Acceleration of oxidative protein folding by newly-developed redox molecules  [Not invited]
    松崎元紀; 岡田隼輔; 荒井堅太; 日高雄二; 稲葉謙次; 村岡貴博; 奥村正樹
    第92回 日本生化学会大会  2019/09
  • メダカ由来プロスタグランジン結合蛋白質の機能解明  [Not invited]
    岩壁一樹; 鳥居希美; 島本 茂; 日高雄二
    第51回若手ペプチド夏の勉強会  2019/08
  • ペプチドホルモン前駆体タンパク質の生理活性構造形成機構  [Not invited]
    大角; 泰唯; 島本 茂; 日高雄二
    第51回若手ペプチド夏の勉強会  2019/08
  • カイココクナーゼの酵素活性化機構の解明  [Not invited]
    竹川 舞; 島本 茂; 日高雄二
    第51回若手ペプチド夏の勉強会  2019/08
  • de novo デザイン蛋白質の立体構造形成機構の解明（ポスター賞受賞）  [Not invited]
    福辻真由; 島本 茂; 日高雄二
    第51回若手ペプチド夏の勉強会  2019/08
  • タンパク質へのSeCys残基の選択的導入の検討  [Not invited]
    藤田勝也; 島本 茂; 日高雄二
    第51回若手ペプチド夏の勉強会  2019/08
  • 心房性ナトリウム利尿ペプチド前駆体タンパク質の構造制御および機能解析  [Not invited]
    上田 隼; 島本 茂; 日高雄二
    第５１回若手ペプチド夏の勉強会  2019/08
  • 分子進化におけるプロウログアニリンの立体構造形成機構の解明  [Not invited]
    マハラジャン アマン ラル; 島本 茂; 日高雄二
    第５１回若手ペプチド夏の勉強会  2019/08
  • 心房性ナトリウム利尿ペプチド前駆体の構造制御及び機能解析  [Not invited]
    上田 隼; 島本 茂; 日高 雄二
    第19回日本蛋白質科学会年会  2019/06
  • ジョロウグモ由来消化酵素のクローニングおよび機能解析  [Not invited]
    田川 翼; 萩原 央記; 宮澤 光博; 島本 茂; 日高 雄二
    第19回日本蛋白質科学会年会  2019/06
  • PDI ファミリーによる前駆体タンパク質フォールディング中間体の分子認識機構の解明  [Not invited]
    増田 光紀; 金村 進吾; 木下 岬; 山口 宏; 日高 雄二; 稲葉 謙次; 奥村 正樹
    第19回日本蛋白質科学会年会  2019/06
  • 造血器型プロスタグランジンD 合成酵素と補酵素・基質の分子認識機構と反応機構の解明  [Not invited]
    東野 貴大; 島本 茂; 日高 雄二
    第19回日本蛋白質科学会年会  2019/06
  • メダカ由来プロスタグランジンD 合成酵素類縁体の機能解明  [Not invited]
    岩壁 一樹; 鳥居 希美; 島本 茂; 加川 尚; 日高 雄二
    第19回日本蛋白質科学会年会  2019/06
  • 等温滴定型熱測定による睡眠物質合成酵素と基質の相互作用解析  [Not invited]
    島本 茂; 中川 悠介; 日高 雄二; 丸野 孝浩; 小林 祐次; 河原 一樹; 吉田 卓也; 大久保 忠恭; 有竹 浩介; 裏出 良博
    第19回日本蛋白質科学会年会  2019/06
  • 新規レドックス分子による酸化的フォールディングの促進と応用  [Not invited]
    松崎 元紀; 岡田 隼輔; 荒井 堅太; 日高 雄二; 稲葉 謙次; 村岡 貴博; 奥村 正樹
    第19回日本蛋白質科学会年会  2019/06
  • 分子進化におけるプロウログアニリンの立体構造形成機構の解明  [Not invited]
    マハラジャン アマン ラル; 島本 茂; 日高雄二
    第35回関西ペプチドセミナー  2018/12
  • 蛋白質へのSeCys残基の選択的導入の検討(ポスター賞受賞）  [Not invited]
    藤田克也; 島本 茂; 日高雄二
    第35回関西ペプチドセミナー  2018/12
  • ジョロウグモ由来酵素のクローニングおよび機能解析(ポスター賞受賞）  [Not invited]
    田川 翼; 島本 茂; 日高雄二
    第35回関西ペプチドセミナー  2018/12
  • 心房性ナトリウム利尿ペプチ前駆体の構造制御および機能解析  [Not invited]
    上田 隼; 島本茂; 日高雄二
    第35回関西ペプチドセミナー  2018/12
  • beta-アミロイド前駆体蛋白質N末端ドメインの構造解析(ポスター賞受賞）  [Not invited]
    今村瑞穂; 島本 茂; 日高雄二
    第35回関西ペプチドセミナー  2018/12
  • l-PGDSの分子進化：メダカL-PGDSの基質認識機構  [Not invited]
    鳥居希美; 島本 茂; 日高雄二
    第35回関西ペプチドセミナー  2018/12
  • Cloning and Functional Analysis of Digestive Enzyme Derived from N ephila Clavata  [Not invited]
    Tsubasa Tagawa; Teruki Hagiwara; Shigeru Shimamoto; Yuji Hidaka
    10th International Peptide Symposium  2018/12
  • Structural Control and Functional Analysis of the Precursor Protein of Atrial Natriuretic Peptide  [Not invited]
    HayatoUeda; Shigeru Shimamoto; Yuji Hidaka
    10th International Peptide Symposium  2018/12
  • Structural Analyses of an N-terminal Extracellular Domain of the Amyloid Precursor Protein  [Not invited]
    Mizuho Imamura; Shingo Kanemura; Masaki Okumura; Hiroshi Yamaguchi; Shigeru Shimamoto; Yuji Hidaka
    10th International Peptide Symposium  2018/12
  • ⼼房性利尿ペプチド前駆体の構造制御と機能に関する研究  [Not invited]
    上田隼; 島本茂; 日髙雄二
    第50回若手ペプチド夏の勉強会  2018/08
  • ジョロウグモ由来消化酵素のクローニングおよび機能解析(ポスター賞受賞）  [Not invited]
    田川翼; 島本茂; 日髙雄二
    第50回若手ペプチド夏の勉強会  2018/08
  • L-PGDS の分⼦進化：メダカL-PGDS の基質認識機構  [Not invited]
    鳥居希美; 島本茂; 日髙雄二
    第50回若手ペプチド夏の勉強会  2018/08
  • β‐アミロイド前駆体蛋⽩質のリンカー領域の構造解析  [Not invited]
    今村瑞穂; 島本茂; 日髙雄二
    第50回若手ペプチド夏の勉強会  2018/08
  • 前駆体タンパク質のプロ領域が制御するフォールディング機構の解明  [Not invited]
    小林 優真; 奥村 正樹; 島本 茂; 稲葉 謙次; 山口 宏; 日高 雄二
    第18回日本蛋白質科学会年会  2018/06
  • MOLECULAR RECOGNITION MECHANISM OF HEMATOPOIETIC PROSTAGLANDIND SYNTHASE WITH COFACTOR AND ITS SUBSTRATE  [Not invited]
    Shigeru Shimamoto; Keisuke Asada; Yuji Hidaka
    62th Annual Meeting of Biophysycal Society  2018/02
  • ACTIVATION MECHANISM OF COCOONASE  [Not invited]
    Nagisa Tajima; Mitsuhiro Miyazawa; Shigeru Shimamoto; Yuji Hidaka
    62th Annual Meeting of Biophysycal Society  2018/02
  • STRUCTURAL ANALYSES OF A LINKER REGION OF THE AMYLOID PRECURSOR PROTEIN  [Not invited]
    Mizuho Imamura; Shingo Kanemura; Masaki Okumura; Shigeru Shimamoto; Yuji Hidaka
    62th Annual Meeting of Biophysycal Society  2018/02
  • FOLDING ANALYSES OF A DE NOVO DESIGNED PROUROGUANYLIN.  [Not invited]
    Yuji Hidaka; Saya Nishihara; Kenta Mori; Shigeru Shimamoto
    62th Annual Meeting of Biophysical Society  2018/02
  • DISULFIDE-COUPLED FOLDING OF PROUROGUANYLIN ON MOLECULAR EVOLUTION  [Not invited]
    Kenta Mori; Saya Nishihara; Shigeru Shimamoto; Yuji Hidaka
    62th Annual Meeting of Biophysical Society  2018/02
  • MOLECULAR EVOLUTION OF L-PGDS: SUBSTRATE RECOGNITION MECHANISM  [Not invited]
    Kimi Torii; Yuji Hidaka; Shigeru Shimamoto
    62th Annual Meeting of Biophysical Society  2018/02
  • アミロイドβ前駆体蛋白質のリンカー領域の立体構造解析(ポスター賞受賞）  [Not invited]
    今村瑞穂; 日高雄二
    第34回関西ペプチドセミナー  2017/12
  • L-PGDSの分子進化：メダカL-PGDSの基質認識機構  [Not invited]
    鳥居希美; 島本 茂; 日高雄二
    第34回関西ペプチドセミナー  2017/12
  • de novo設計蛋白質の立体構造形成機構の解明(ポスター賞受賞）  [Not invited]
    西原 紗彩; 日高雄二
    第34回関西ペプチドセミナー  2017/12
  • プロペプチドによるカイココクナーゼの立体構造形成機構の解明  [Not invited]
    田嶋 渚; 日高雄二
    第34回関西ペプチドセミナー  2017/12
  • 分子進化におけるプロウログアニリンの立体構造形成機構の解明  [Not invited]
    森 健太; 日高雄二
    第34回関西ペプチドセミナー  2017/12
  • Disulfide-coupled folding of pro-uroguanylin on molecular evolution  [Not invited]
    Kenta Mori; Kosuke Toyama; Saya Nishihara; Shigeru Shimamoto; Yuji Hidaka
    The 54th Japanese Peptide Symposium  2017/11
  • Molecular evolution of L-PGDS: substrate recognittion mechanism of medaka L-PGDS  [Not invited]
    Kimi Torii; Takahiro Maruno; Yuji Kobayashi; Yuji Hidaka; Shigeru Shimamoto
    The 54th Japanese Peptide Symposium  2017/11
  • Regulation of disulfide-coupled folding of a de novo designed protein  [Not invited]
    Saya Nishihara; Kosuke Toyama; Kenta Mori; Shigeru Shimamoto; Yuji Hidaka
    The 54th Japanese Peptide Symposium  2017/11
  • Structural analyses of a linker region of the amyloid precursor protein  [Not invited]
    Mizuho Imamura; Shingo Kanemura; Masaki Okumura; Shigeru Shimamoto; Yuji Hidaka
    The 54th Japanese Peptide Symposium  2017/11
  • Lipocalin-Type Prostaglandin D Synthase Possesses Two Binding Sites for its product.  [Not invited]
    Shigeru Shimamoto; Yusuke Nakagawa; Yuta Nakahata; Takahiro Maruno; Yuji Kobayashi; Kosuke Aritake; Yoshihiro Urade; Yuji Hidaka
    61st Annual Meeting of Biophysical Scoiety  2017/02
  • Molecular recognition mechanism of Hematopoietic Prostaglandin D Synthase with cofactor and its substrate  [Not invited]
    Keisuke Asada; Shigeru Shimamoto; Tomohiro Oonoki; Takahiro Maruno; Yuji Kobayashi; Kosuke Aritake; Yoshihiro Urade; Yuji Hidaka
    61st Annual Meeting of Biophysical Scoiety  2017/02
  • Regulation of protein folding using organic solvents and ion liquid  [Not invited]
    Yuji Hidaka; Wataru Ito; Ryosuke, Nishimura; Shigeru Shimamoto
    61st Annual Meeting of Biophysical Scoiety  2017/02
  • Structural analysis of the precursor protein of atrial natriuretic peptide  [Not invited]
    Sumika Futori; Satomi Higashigawa; Shigeru Shimamoto; Yuji Hidaka
    61st Annual Meeting of Biophysical Scoiety  2017/02
  • Regulation of folding of de novo designed peptides by alpha-helix formation  [Not invited]
    Saya Nishihara; Kosuke Toyama; Shigeru Shimamoto; Yuji Hidaka
    61st Annual Meeting of Biophysical Scoiety  2017/02
  • Preparation of a orexin precursor protein by chemical digestion  [Not invited]
    Natsumi Mitsuoka; Shigeru Shimamoto; Yuji Hidaka
    61st Annual Meeting of Biophysical Society  2017/02
  • de novo設計蛋白質の立体構造形成機構の解明  [Not invited]
    西原 紗彩; 島本 茂; 日高雄二
    第33回 関西地区ペプチドセミナー  2016/11
  • 分子進化におけるプロウログアニリンの立体構造形成機構の解明  [Not invited]
    森 健太; 島本 茂; 日高雄二
    第33回 関西地区ペプチドセミナー  2016/11
  • プロペプチドによるカイココクナーゼの立体構造形成  [Not invited]
    田嶋 渚; 島本 茂; 日高雄二
    第33回 関西地区ペプチドセミナー  2016/11
  • 化学切断法による酵素分解性遺伝子組換え蛋白質の調製(ポスター最優秀賞受賞）  [Not invited]
    光岡夏海; 島本 茂; 日高雄二
    第33回 関西地区ペプチドセミナー  2016/11
  • 造血型プロスタグランジン D 合成酵素と補酵素･基質の分子認識機構と反応機構の解明  [Not invited]
    浅田恵佑; 島本 茂; 日高雄二
    第33回 関西地区ペプチドセミナー  2016/11
  • DISULFIDE-COUPLED FOLDING OF PRO-UROGUANYLIN ON MOLECULAR EVOLUTION  [Not invited]
    Kenta Mori; Kosuke Toyama; Shigeru Shimamoto; Yuji Hidaka
    10th International Peptide Symposium  2016/10
  • ACTIVATION MECHANISM OF COCOONASE  [Not invited]
    Nagisa Tajima; Shigeru Shimamoto; Mitsuhiro Miyazawa; Yuji Hidaka
    10th International Peptide Symposium  2016/10
  • REGULATION OF DISULFIDE-COUPLED FOLDING OF DE NOVO DESIGNED PEPTIDES BY alpha-HELIX FORMATION  [Not invited]
    Saya Nishihara; Kosuke Toyama; Shigeru Shimamoto; Yuji Hidaka
    10th International Peptide Symposium  2016/10
  • PREPARATION OF A OREXIN PRECORSOR PROTEIN USING E. COLI EXPRESSION SYSTEM BY ACID TREATMENT  [Not invited]
    Natsumi Mitsuoka; Shigeru Shimamoto; Yuji Hidaka
    10th International Peptide Symposium  2016/10

MISC

Research Grants & Projects

  • 日本学術振興会：科学研究費(基盤研究Ｃ）
    Date (from‐to) : 2016/04 -2020/03 
    Author : 日高雄二
  • ペプチドホルモン前駆体蛋白質を標的とした分子進化と生理活性成熟化機構の解明
    日本学術振興会：科学研究費(基盤研究Ｃ）
    Date (from‐to) : 2012/04 -2016/03 
    Author : 日高雄二
  • 脱化石原料のための分子改変による植物由来有機合成原料の創出
    科学技術振興機構：A L C A 研究開発
    Date (from‐to) : 2011/10 -2012/09 
    Author : 日高雄二
  • プロペプチドの分子内シャペロン機能の解明と新規生理活性ペプチドの創作
    日本学術振興会：科学研究費(基盤研究Ｃ）
    Date (from‐to) : 2008/04 -2012/03 
    Author : 日高雄二
  • 前駆体タンパク質の構造生物学的研究を基礎とした生理活性ペプチド成熟機構の解明
    日本学術振興会：科学研究費(基盤研究Ｃ）
    Date (from‐to) : 2007/04 -2010/03 
    Author : 山口 宏
  • プロペプチドの分子内シャペロン機能を利用した新規生理活性ペプチドの構築
    日本学術振興会：科学研究費(基盤研究Ｃ）
    Date (from‐to) : 2004/04 -2008/03 
    Author : 日高雄二
  • 腸管上皮細胞膜結合型グアニル酸シクラーゼ活性化ペプチドのシグナル伝達機構の解明
    日本学術振興会：科学研究費(奨励研究Ａ）
    Date (from‐to) : 1993/04 -1994/03 
    Author : 日高雄二

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