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Research on the structure required for function of protease inhibitor proteins produced by bacteria by protein engineering

Research Project

Project/Area Number 02305003
Research Category

Grant-in-Aid for Co-operative Research (A)

Allocation TypeSingle-year Grants
Research Field 広領域
Research InstitutionGAKUSHUIN UNIVERSITY, FACULTY OF SCIENCE(INSTITUTE FOR BIOMOLECULAR SCIENCE) (1991-1992)
The University of Tokyo (1990)

Principal Investigator

MIURA Kin-ichiro  GAKUSHUIN UNIVERSITY PROFESSOR, 理学部, 教授 (30000227)

Co-Investigator(Kenkyū-buntansha) AKASAKA Kazuyuki  KOBE UNIVERSITY PROFESSOR, 理学部, 教授 (50025368)
TONOMURA Ben-ichiro  KYOTO UNIVERSITY PROFESSOR, 農学部, 教授 (20026545)
KAINOSHO Masatsune  TOKYO METROPOLITAN UNIVERSITY PROFESSOR, 理学部, 教授 (20137029)
OHTA Takahisa  UNIVERSITY OF TOKYO PROFESSOR, 農学部, 教授 (30011844)
MITSUI Yukio  NAGAOKA TECHNOLOGY and SCIENCE UNIVERSITY PROFESSOR, 工学部, 教授 (40012637)
Project Period (FY) 1990 – 1992
Project Status Completed (Fiscal Year 1992)
Budget Amount *help
¥11,200,000 (Direct Cost: ¥11,200,000)
Fiscal Year 1992: ¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 1991: ¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 1990: ¥6,300,000 (Direct Cost: ¥6,300,000)
Keywordsprotein engineering / enzyme inhibitor / protease inhibitor / site-directed mutagenesis / bacteria / gene engineering / functional site in a protein / protein structure / 阜ロテイーゼ・インヒビター / プロテア-ゼ・インヒビタ-
Research Abstract

The structure required for an inhibitor against a protease was studied by protein engineering with the use of protease inhibitor proteins produced by bacteria, mainly Streptomyces subtilisin inhibitor (SSI).The center of reactive site of SSI (methionine) was replaced by other 19 kinds of amino acid. Mutants replaced by a basic amino acid (lysine or arginine) tended to inhibit trypsin that splits a protein at the positions of basic amino acid residues, although the wild type SSI inhibits trypsin scarcely. Only the lysine mutant inhibited lysyl endopeptidase. The mutant SSIs replaced by an aromatic amino acid (tryptophane, tyrosine, and phenylalanine) tended to inhibit chymotrypsin, which splits a protein at the aromatic amino acid residues. Single amino acid change caused alteration of the functional specificity of a protein. Thus, the reactive site in a protease inhibitor should be similar to a substrate structure to fit to the active center of a protease.
The reactive site is supported by other parts of SSI molecule. For example, if the S-S bridge near the reactive site was taken off by replacing two cysteine residues by serine residues, SSI tended to a substrate for proteases. From these experiments we reached to the conclusion that the reactive site in an inhibitor protein molecule should be supported firmly.
The study of physical properties of the similar protease inhibitors, which are obtained not only from bacteria but also fungi and plant seeds, and of the artificially modified inhibitors was carried out. The interaction of these inhibitors and a protease was also studied by X-ray analysis, NMR, and other spectral analysis. From these results the following conclusion was obtained: A fine structural change is caused easily in the reactive site and the part near C-terminal, so that the reactive site causes induced fitting on the interaction with the active site of a protease.

Report

(4 results)
  • 1992 Annual Research Report   Final Research Report Summary
  • 1991 Annual Research Report
  • 1990 Annual Research Report
  • Research Products

    (97 results)

All Other

All Publications (97 results)

  • [Publications] H.Takahashi,T.igarashi,I.Shimada and Y.Arata: "Preparation of the Fragment from a Short-Chain Mouse lgG2a Anti-Dansyl Monoclonal Antibody and Use of Selectively Deuterated Fv Analogues for Two Dimensional ^1H NMR Analysis of the Antigen-Antibody lnteractions" Biochmistry. 30. 2840-2847 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] K.Kato,C.Matsunaga,A.Odaka,S.Yamato,W.Takaha,I.Shimada and Y.Arata: "Carbon-13 NMR Study of Switch Variant Anti Dansyl Antibodies:Antigen Binding and Domain-Domain lnteractions" Biochemistry. 30. 6604-6610 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] C.Matsunaga,K.Kato and Y.Arata: "A ^<13>CNMR study of the hinge region of a mouse monoclonal antibody" J.Biomolecular NMR. 1. 379-390 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] J.I.Kim,T.Nagano,T.Higuchi,M.Hirobe,I.Shimada and Y.Arata: "Conformation and Stereoselective Reduction of Hapten Side Chains in the Antibody Combining Site" J.American Chemical Society. 113. 9392-9394 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] H.Takahashi,E.Suzuki,K.Shimada and Y.Arata: "Dynamical Stucture of the Antibody combinig Site As Studied ^1H- ^<15>N Shift Correlation NMR Spectroscopy" Biochemistry. 31. 2464-2468 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] H.Gouda,H.Torigoe,A.Saito,M.Sato,Y.Arata and I.Shimada: "Three-Dimensional Solution STructure of the B Domain of Staphylococcal Protein A:Comparisons of the Solution and Crystal Structures" Biochemistry. 31. 9665-9672 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] A.Odaka,J.I.Kim,H.Takahashi,I.Shimada and Y.Arata: "lsotope Edited Nuclear Magnetic Resonace Study of Fv Fragment of Anti-Dansyl Mouse Monoclonal Antibody:Recognition of the Dansyl Hapten" Biochimistry. 31. 10686-10691 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] K.Uchida,Y.Miyake and M.Kainosho: "Reactive Cleavage and recognition of the disulfie bonds in Strptmyces subutilisin inhibitor(SSI) as Studied by the carbonyl ^<13>C NMR resonances of cycteinyl residues" J.of Biomolecular NMR. 1. 49-64 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] K.Uchida and M.Kainosho: "Synthesis of [1,1'- ^<13>C_2]-L-Cystine" J.Labelled Compounds and Radiopharmaceuticals. XXIX. 867-874 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] 甲斐荘 正恒,安倍 未央,三宅 洋子,小島 修一: "蛋白質の安定同位体利用NMRスペクトル" 蛋白質・核酸・酸素. 37. 269-283 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Y.Takeuchi,Y.SatoW,K.T.Nakamura and Y.Mitsui: "Refined Crystal Structure of the Comlex of Subtilisin BPN' and Streptomyces Subtilisin Inhibitor at 1.8A Resolution" J.Molecular Biology. 221. 309-325 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] V.Takeuchi,S.Noguchi,Y.Satow,S.Kojima,I.Kumagai,K.Miura,K.T.Nakamura and Y.Mitsui: "Molecular recognition at the active site of subtilisin BPN':crystallographic studies using genetically engineered proteinaceous inhibitor SSI(Streptomyces subutilisin inhibitor)" Protein Engineering. 4. 501-508 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] Y.Takeuchi,T.Nonaka,K.T.Nakamura,S.Kojima,K.Miura and Y.Mitsui: "Crystal structure of an engineered subtilisin inhibitor complexed with bovine trypsin" Proc.Natl.Acad.Sci.USA. 89. 4407-4411 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] A.Tamura,K.Kanamori,S.Kojima,I.Kumagai,K.Miura and K.Akasaka: "Mechanisms of Temporary Inhibition is Streptomyces Subtilisin Inhibitor Induced by an Amino Acid Substitution,Tryptophan 86 Replaced by Histidine" Biochemistry. 30. 5275-5286 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] A.Tamura,K.Kimura,H.Takahara and K.Akasaka: "Cold Denaturation and Heat Denaturation of Streptomyces Subtilisin Inhibitor 1.CD andDSC Studies" Biochimistry. 30. 11307-11313 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] A.Tamura,K.Kimura and K.Akasaka: "Cold Denaturation and Heat Denaturation of Streptomyces Subtilisin Inhibitor 2. ^1H NMR Studies" Biochemistry. 30. 11313-11320 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] I.Terada,S.T.Kwon,Y.Miyata,H.Matsuzawa and T.Ohta: "Unique Precursor Structure of an Extracellular Protease,Aqualysin I,with NH_2-and COOH-treminal Pro-sequences and its Processing in Escherichia coli" J.Biological Chemistry. 265. 6576-6581 (1990)

    • Related Report
      1992 Annual Research Report
  • [Publications] N.Touhara,H.Taguchi,Y.Koyama,T.Ohta and H.Matsuzawa: "Production and Extracellular Secretion of Aqualysin I(a Thermophilic Subtilisin-Type Protease)in a Host-Vector System for Thermus thermophilus" Applied and Environment Microbiology. 57. 3385-3387 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] Y.C.Lee,Y.Miyata,I.Terada,T.Ohta and H.Matsuzawa: "Involvement of NH_2-Terminal pro-sequence in the production of Active Aqualysin I(a Thermophlic Serine Protease)in Escherichia coli" Agricultural and Biological Chemistry. 55. 3027-3032 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] Y.C.Lee,T.Ohta and H.Matsuzawa: "A non-covalent NH_2-terminal pro-region aids the production of active aqualysin I(a thermophilic protease)without the COOH-terminal pro-sequence in Escherichia coli" FEMS Microbiology Letters. 92. 73-78 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] K.Miura,I.Kumagai,S.Obata,S.Kojima and S.Taguchi: "Partial Alteration of a Protein Streptomyces Subutilisin Inhibitor by Site-Directed Mutagenesis" Proc.Japan Acad.64SerB. 147-149 (1988)

    • Related Report
      1992 Annual Research Report
  • [Publications] S.Obata,S.Taguchi,I.Kumagai and K.Miura: "Molecular Cloning and Nucleotide Sequence Determination of Gene Encodiong Streptomyces Subtilisin Inhibitor(SSI)" J.Biochemistry. 105. 367-371 (1989)

    • Related Report
      1992 Annual Research Report
  • [Publications] S.Obata,S.Furukubo,I.Kumagai,H.Takahasi and K.miura: "High-Level Expression in Streptomyces Lividans 66 of a Gene Encoding Streptomyces Subutilisin Lnhibitor from Stretomyces albogriseolus S-3253" J.Biochemistry. 105. 372-376 (1989)

    • Related Report
      1992 Annual Research Report
  • [Publications] S.TAguchi,K.Nisiyama,I.Kumagai and K.Miura: "Analysis of transcriptional control regions in the Streptomyces subtilisin-inhibitor-encoding gene" Gene. 84. 279-286 (1989)

    • Related Report
      1992 Annual Research Report
  • [Publications] S.Taguchi,I.Kumagai,J.Nakayama,A.Suzuki and K.Miura: "EFFICIENT EXTRACELLULAR EXPRESSION OF A FOREIGN PROTEIN IN STREOTOMYCES USUNG SECRETORY PROTEASE INHIBITOR(SSI) GENE FUSIONS" Bio/technology. 7. 1063-1066 (1989)

    • Related Report
      1992 Annual Research Report
  • [Publications] S.Kojima,I.Kumagai and K.Miura: "Effect on inhibitor activity of mutation at reactionasite P4 of the Streptpmyces subtilisin inhibitor,SSI" Protein Engineering. 3. 527-530 (1990)

    • Related Report
      1992 Annual Research Report
  • [Publications] S.Kojima,S.Obata,I.Kumagai and K.miura: "ALTERATION OF THE SPECIFICITY OF THE STREOTOMYCES SUBUTILISIN INHIBITOR BY GENE ENGINEERING" Bio/technology. 8. 449-452 (1990)

    • Related Report
      1992 Annual Research Report
  • [Publications] S.Taguchi,I.Kamagai and K.Miura: "Comparison of seretory expression in Escherichia coli and Streptomyces of Streptomyces subtilisin inhibitor(SSI) gene" Biochimica et Biophysica Acta. 1049. 278-285 (1990)

    • Related Report
      1992 Annual Research Report
  • [Publications] S.Kojima,Y.Nishiyama,I.Kumagai K.Miura: "Inhibition of Subtilisin BPN' by Reaction Site P1 Mutants of Strepromyces Subtilisin Inhibitor" J.Biochimistry. 109. 377-382 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] S.Taguchi,K.Nishiyama,I.Kumagai and Miura: "Relationship between utilization of dual translational initiation signals and protein procession in Streptomyces" Mol.Gen.Genet.226. 328-331 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] Y.Ueda,S.Kojima,K.Tsumoto,S,Takeda,K.Miura and I.Kumagai: "A Protease Inhibitor Produced by Streptomyces lividans 66 Exhibits Inhibitory Activities toward Both Subtilisin BPN' and Trypsin" J.Biochemistry. 112. 204-211 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] S.TAguchi,S.Kojima,I.Kumagai,H.Ogawara,K.Miura and H.Momose: "Isolation and partial characterization of SSI-like protease inhibitors from Streptomyces" FEMS Microbiology Letters. 99. 293-298 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] S.Kojima,I.Kumagai and K Miura: "Requirement of Disulfide Bridge Near the Reactive Site of Protease Inhibitor SSI(Streptomyces Subtilisin Inhibitor) for Its Inhibitory" J.Molecular Biology. 230. (1993)

    • Related Report
      1992 Annual Research Report
  • [Publications] S.Taguchi,H,Kikuchi,S.Komima,I.Kumagai,T.Nakase,K.Miura and H.Momose: "High Frequency of SSI-like Protease Inhibitors among Streptomyces" Biosci.Biotech.Biochem.57. 522-524 (1993)

    • Related Report
      1992 Annual Research Report
  • [Publications] K.M-Monnma,T.Hatanaka,K.Inouye,K.Kanamori,A.Tamura,K.Akasaka,S.Komima,K.Miura and B.Tonomura: "Interaction of Subtilisin BPN' and Recombinant Streptomyces Subtilisin Inhibitors of Which the P_1 Site Residue in Replaced" J.Biochemistry.

    • Related Report
      1992 Annual Research Report
  • [Publications] T.Kumazaki,N.Hoshiba,H.Yokosawa and S.Ishii: "Primary Structure of Ascidian Trypsin Inhibitors in the Hemolymph of a Solitary Ascidian,Halocynthia roretzi" J.Biochemistry. 107. 409-413 (1990)

    • Related Report
      1992 Annual Research Report
  • [Publications] T.Kumazaki and S.Ishii: "Disulfide Bridge Structure of Ascidian Trypsin Inhibitor I:Similarity to Kazal-Type Inhibitors" J.Biochemistry. 107. 414-419 (1990)

    • Related Report
      1992 Annual Research Report
  • [Publications] H.Tsuyaki,K.Kamiwara,A.Fujita,T.Kumazaki and S.Ishii: "Purification and Characterization of Streptomyces griseus Metalloendopeptidase I and II" J.Biochemistry. 110. 339-344 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] K.Kamiwara,T.Kumazaki,E.Sato,Y.Kanaoka and S.Ishii: "Application of Bimane Peptide Substrate to Spectrofluorometric Assays of Metalloendopeptidase" J.Biochemistry. 110. 345-349 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] K.Kajiwara,A.Fujita,H.Tsuyaki,T.Kumazaki and S.Ishii: "Interaction of Streptomyces Serine Protease Inhibitors with Streptomyces Griseus Metalloendopeptidase II" J.Biochemistry. 110. 35-354 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] K.Tanaka,H.Saito,M.Arai,S.Murao and H.Takahashi: "Cloning and Expression of Metallo-prpteinase Inhibitor(S-MP1) Gene From Streptomyces nigrescens" Biochemical and Biophysical Research Communications. 155. 487-492 (1988)

    • Related Report
      1992 Annual Research Report
  • [Publications] T.Shiina,K,Tanaka and H.Takahasi: "Sequence of hrdB,an essential gene encoding sigma-like transcription factor of Streptomyces coelicolor A3(2): homology to principal sigma factors" Gene. 107. 145-148 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] K.Tanaka,T.Shiina and H.Takahashi: "Nucleotide sequence of gene hrdA,hrdC,and hrdD for Streptomyces coelicolor A3(2) having similarity to rpoD genes" Mol.Gen.Genet.229. 334-340 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] K.Tanaka,H.Aoki,K.Oda,S.Murao.H.Saito and H.Takahashi: "Mucleotide sequence of the gene for a metallopoteinase inhibitor of Streptomyces nigrescens(SMPI)" Nucleic Acid Research. 18. 6433-6433 (1990)

    • Related Report
      1992 Annual Research Report
  • [Publications] K.Oda,H.Nakatani and B.M.Dunn: "Substrate specificity and kinetic properties of pepstatin-intensive carboxly proteinase from Pseudomonas sp.No.101" Biochimica et Biophysica Acta. 1120. 208-214 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] S.Murao,K.Ohkuni,M.Nagao,K.Oda and T.Shin: "A novel Thermostable,S-PI(Pepstatin Ac)-insensitive Acid Proteinase from Thermophilic Bacillus Novosp.Starain MN-32" Agricultural and Biological Chemistry. 52. 1629-1631 (1988)

    • Related Report
      1992 Annual Research Report
  • [Publications] K.Oda.Y.Fukuda,S.Murao,K.Uchida and M.Kainosho: "A Novel Proteinase Inhibitor,Inhibiting Some Pepstatine-insensetive Carboxy Proteinases" Agricultural and Biological Chemistry. 53. 405-415 (1989)

    • Related Report
      1992 Annual Research Report
  • [Publications] K.Oda and S.Murao: "PEPSTATIN INSENSITIVE CARBOXYL PROTEINASES" Stucture and Function of the Aspartic proteinases. 185-201 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] S.Murao,K.Ohkuni,M.Nagao,K.hirayama,K.Fukuhara,K.Oda,H.Oyamaand T.Shin: "Purification and Characterization of Kumamolysin,a Novel Thermostable Pepstatin-insensitive Carboxyl Proteinase from Bacillus novosp.MN-32" J.Biological Chemistry. 268. 349-355 (1993)

    • Related Report
      1992 Annual Research Report
  • [Publications] D.Tsuru,T.Yoshimoto,N.Koriyama and S.Furukawa: "Thiazolidine Derivatives as Potent Inhibitors Specific for Prolyl Endopeptidase" J.Biochemistry. 104. 580-586 (1988)

    • Related Report
      1992 Annual Research Report
  • [Publications] R.Kobayashi,T.Yoshimoto and D Tsuru: "Coplete Amino Acid Sequence of Neutral Protease from Bacillus subtilis var.amylosacchariticus" Agricultural and Biological Chemistry. 53. 2737-2749 (1989)

    • Related Report
      1992 Annual Research Report
  • [Publications] R.Kobayashi,A Kanatani,T.Yasimoto and D.Tsuru: "Chemical Modification of Neutral Protease from Bacillus subtilis var.amylosacchariticus with Tetranitromethane:Assignment of Tyrosyl Residues Nitrated" J.Biochemistry. 106. 110-113 (1989)

    • Related Report
      1992 Annual Research Report
  • [Publications] T.Yoshimoto,H.Oyama,T.Takeshita,H.Higashi,S.L.Xu and D.Tsuru: "Nucleotide Sequence of the Neutral Protease Gene from Bacillus subtilis var.amylosacchariticus" J.Fermentation And Bioengineering. 70. 370-375 (1990)

    • Related Report
      1992 Annual Research Report
  • [Publications] T.Yoshimoto,D.Tsuru,N.Yamamoto,R.lkezawa and S.Furukawa: "Structure Activity Relationship of Inhibitors Specific for Prolyl Endopeptidase" Agriultural and Biological Chemistry. 55. 37-43 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] S.Murao,Y.Nomura,K.Nagamatu,K.Hirayama,M.Iwahara and T.Shin: "Purification and Some Properties of a Thermostable Metal Proteinase Produced by Thermomicrobium sp.KN-22 Strain" Agricultural and Bilogical Chemistry. 55. 1739-1744 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] S.Morikawa,A.Kanatani,T.Yoshimoto and D.Tsuru: "Dye-sensitized photooxidation of Neutral Protease from Bacillus subtilis var.amylosacchariticus:Assginment of Histidine Residues Oxdized" Agricultural and Bilogical Chemistry. 55. 2099-2103 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] A.Kanatani,T.Matuda,T.Shimoda,F.Misoka,X.S.Lin,T.Yoshimoto and D.Tsuru: "ProteaseII from Escherichia coli:Sequencing and Expression of the Enzyme Gene and Charactrization of the Expressed Enzyme" J.Biochemistry. 110. 315-320 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] T.Yoshimoto,A.Kanatami,T.Shimoda,T.Inaoka,T.Kokubo and D.Tsuru: "Prolyl Endopeptidase from Flavobactreium meningosepticum:Cloninng and Sequencimg of the Enzyme Gene" J.Biochemistry. 110. 873-878 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] S.Morikawa,A.Kanatami,R.T.Yoshimoto and D.Tsuru: "Chemical Modification of Neutral Protease from Bacillus subtilis var.amylosacchariticus:Assignment of Tyrosyl Residues lodinated" Agricultural and Biological Chemistry. 55. 2751-2756 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] D.Tsuru,Y.Fujita,S.Morikawa,K.Ito and T.Yoshimoto: "Degradation of Streptpmyces Metalloprotease Inhibitor(SMPI) by Neutral Protease from Bacillus subtilis var.amylosacchariticus" Biosci.Biotech.Biochem. 56. 1275-1278 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] A.Nakatani,T.Yosimoto H.Nagai,K.Ito and D.Tsuru: "Location of the Protease II Gene (prtB) on the Physical Map of the Escherichia coli Chromosome" J.Bacteriology. 174. 7881-7881 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] T.Yosimoto,T.Shimoda,A.Kitazono,T.Kabashima,K.Ito and D.Tsuru: "Pyroglutamyl Peptidase Gene from Bacillus amyloliquefaciens:Cloning,Sequencing,Expression,and Crystallization of the Expressed Enzyme" J.Biochemistry. 113. 67-73 (1993)

    • Related Report
      1992 Annual Research Report
  • [Publications] R.Takano,C.Imada,K.Kamei and S.Hara: "The Reactive Site of Marinostatin,a Proteinase Inhibitor from Marine Alteromonas sp.B-10-31" J.Biochemistry. 110. 856-858 (1991)

    • Related Report
      1992 Annual Research Report
  • [Publications] N.Hamato,R.Takano,K.K-Hayashi,C.Imada and S.Hara: "Leupeptins Produced by the Marine Alteromonas sp.B-10-31" Biosci.Biotech.Biochem.56. 1316-1318 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] S.Taguchi,K.Nishiyama,I.Kumagai,H.Momose,K.Miura: "Relationship between ntilization of dual translation initiation signals and protein processing in Streptomyces" Mol.Gen.Genet.226. 328-331 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] S.Kojima,Y.Nishiyama,I.Kumagai,K.Miura: "Inhibition of Subtilisin BPN' by Reaction Site P1 Mutants of Strptmyces Subtilisin Inhibitor" J.Biochem.109. 377-382 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Y.Takeuchi,S.Noguchi,Y.Satow,S.Kojima,I.Kumagai,K.Miura,K.Nakamura,Y.Mitsui: "Molecular recognition at the active site of subtilisin BPN': crystallographic studies using genetically engineered proteinaceous inhibitor SSI(Streptomyces subtilisin inhibitor)" Protein Engineering. 4. 501-508 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] A.Tamura,K.Kanaori,S.Kojima,I.Kumagai,K.Miura,K.Akasaka: "Mechanisms of Temporary Inhibition in Streptomyces Subtilisin Inhibitor Induced by an Anino Acid substitution,Tryptophan 86 Replaced by Histidine" Biochemistry. 30. 5275-5286 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] A.Tamura,K.Kimura,H.Takahara,K.Akasaka: "Cold Denaturation and Heat Denaturation of Streptomyces Subtilisin Inhibitor. 1.CD and DSC Studies" Biochemistry. 30. 11307-11313 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] A.Tamura,K.Kimura,K.Akasaka: "Cold Denaturation and Heat Denaturation of Streptomyces Subtilisin Inhibitor. 2.H NMR Studies" Biochemistry. 30. 11313-11320 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] 三浦 謹一郎,京極 好正,菊池 正和,松浦 良樹 編 三井 幸雄,甲斐荘 正恒,荒田 洋治,三浦 謹一郎 他著: "蛋白質工学の進展ー蛋白質の設計をめざして(蛋白質・核酸・酵素増刊475巻)" 共立出版, 498 (1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] K.Cda,Y.Fukuda,S.Murao K.Uchida and M.Kainosho: "A Novel Proteinase Inhibitor,Inhibiting Some PepstatinーInsensitive Carboxyl Proteinases" Agric.Biol.Chem.53. 405-415 (1989)

    • Related Report
      1990 Annual Research Report
  • [Publications] D.Tsuru,A.Naotuka,R.Kobayashi,T.Yoshimoto,K.Oda and S.Murao: "Inactivation of <Scytalidium>___ー <lignicolum>___ー Acid Protease B with 1,2ーEpoxyー3ー(4′ーdiazoー2′ーnitrophenoxy)propane" Agric.Biol.Chem.53. 2751-2756 (1989)

    • Related Report
      1990 Annual Research Report
  • [Publications] 村尾 沢夫,小田 耕平: "放線菌由来のメタルプロテア-ゼインヒビタ-" 蛋白質・核酸・酵素. 34. 980-988 (1989)

    • Related Report
      1990 Annual Research Report
  • [Publications] S.ーH.Kim,S.Hara,T.Ikenaka: "Reconstitution of new inhibitors by mutual exchange of Ile(64)ーMet(84) peptides of soybean trypsin inhibitors,Ti^a and Ti^b." J.Biochem.

    • Related Report
      1990 Annual Research Report
  • [Publications] R.Takano,C.Imada,K.Kamei,S.Hara: "The reactive site of the novel peptide serine proteinase inhibitor,marinostatin" J.Biochem.

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      1990 Annual Research Report
  • [Publications] T.Yoshimoto,D.Tsuru,N.Yamamoto,R.Ikezawa,S.Furukawa: "Structure activity relationship of inhibitors specific for prolyl endopeptidase." Agricultural Biological Chemistry. 55. 37-43 (1991)

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      1990 Annual Research Report
  • [Publications] K.Akasaka,R.Ishima,and S.Shibata,: "Proton spin relaxation in proteins at high mangetic fields." Physica B. 164,. 163-177 (1990)

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      1990 Annual Research Report
  • [Publications] S.Shibata and K.Akasaka,: "Computer simulation program of proton spin relaxation in proteins and its application to Streptomyces subtilisin inhibitor." Magnetic Resonance in Chemistry. 28,. 129-137 (1990)

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      1990 Annual Research Report
  • [Publications] R.Ishima,S.Shibata,and K.Akasaka,: "General features of proton longitudinal relaxation in proteins in solution." J.Magn.Reson.,in press.91. (1991)

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      1990 Annual Research Report
  • [Publications] S.Murao,K.Ohkuni,T.Shin: "Characterization of thermostable acid proteinase from thermophilic <Bacillus>___ー novosp.MNー32." Agric.Biol.Chem.

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      1990 Annual Research Report
  • [Publications] I.Terada,SukーTae Kwon,Y.Miyata,H.Matsuzawa,T.Ohta: "Unique precursor structure of an extracellular protease,aqualysin I.with NH_2ーand COOHーterminal proーsequences and its processing in <Escherichia>___ー <coli>___ー." J.Biol.Chem.265. 6576-6581 (1990)

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      1990 Annual Research Report
  • [Publications] 加藤,松永,西村,ベルヒリ,甲斐荘,荒田: "Application of ^<13>C nuclear maguetic resonance spectroscopy to molecular structural auslyses of autibody molecules." J.Biochem.105. 867-869 (1989)

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      1990 Annual Research Report
  • [Publications] 小田,福田,村尾,内田,甲斐荘: "A novel proteinase inhibitor,tyrostatin,inhibiting some pepstatinーinsensitime proteinases." Agric.Bilo.Chem.53. 405-415 (1989)

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      1990 Annual Research Report
  • [Publications] 内田,三宅,甲斐荘: "Reductive cleavage and regeneration of the disulfide bonds in <Streptomyces>___ー subtilisin inhibitor (SSI) as studied by the carbonyl ^<13>C NMR resonances of cysteinyl residues." J.Biomol.NMR. 1. 000-000 (1991)

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      1990 Annual Research Report
  • [Publications] 内田,甲斐荘: "Synthesis of 〔1,1′ー^<13>C_2〕ーLーCystive." J.Labelled Compd.Radiopharmacenticals.

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      1990 Annual Research Report
  • [Publications] S.Taguchi,I.Kumagai,K.Miura: "Comparison of secretory expression in Escherichia coli and Streptomyces of streptomyces subtilisin inhibitor (SSI) gene." Biochim.Biophys.Acta. 1049. 278-285 (1990)

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      1990 Annual Research Report
  • [Publications] S.Kojima,I.Kumagai,K.Miura: "Effect on inhibitory activity of mutation at reaction site P4 of tne streptomyces subtilisin inhibitor,SSI." Protein Engineering. 3. 527-530 (1990)

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      1990 Annual Research Report
  • [Publications] I.Kumagai,S.Takeda,T.Hibino,K.Miura: "Expression of goat αーlactalbumin in Escherichia coli and its refolding to biologically active protein." Protein Engineering. 3. 449-452 (1990)

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      1990 Annual Research Report
  • [Publications] S.Kojima,S.Obata,I.Kumagai,K.Miura: "Alteration of the specificity of the streptomyces subtilisin inhibitor by gene engineering." BIO/TECHNOLOGY. 8. 449-452 (1990)

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      1990 Annual Research Report
  • [Publications] S.Taguchi,I.Kumagai,J.Nakayama,A.Suzuki,K.Miura: "Efficient extracellular expression of a foreign protein in Streptomyces using secretory protease inhibitor (SSI) gene fusions." BIO/TECHNOLOGY. 7. 1063-1066 (1989)

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      1990 Annual Research Report
  • [Publications] S.Taguchi,K.Nishiyama,I.Kumagai,K.Miura: "Analysis of transctiptional control regions in the <Streptomyces>___ー subtilisinーinhibitorーencoding gene." Gene. 84. 279-286 (1989)

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      1990 Annual Research Report
  • [Publications] Y.Yoshinaka,I.Katoh and K.Oda: "Retroviral Proteases (ed.L.H.Pearl) Retroviral Protease:Substrate Specificity and Inhibitorsの項執筆" Stockton Press, 9 (1990)

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      1990 Annual Research Report
  • [Publications] K.Oda and S.Murao: "Aspartic Proteinases (ed.B.M.Dunn) Pepstatinーinsensitive Carboxyl Proteinasesの項執筆" Plenum Press, 16 (1991)

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      1990 Annual Research Report
  • [Publications] 鶴 大典: "プロテア-ゼとそのインヒビタ-" メジカルビュ-社, 20 (1991)

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      1990 Annual Research Report
  • [Publications] 甲斐荘,内田(共著): "生物物理のフロンティア" 培風館, 291 (1989)

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      1990 Annual Research Report
  • [Publications] 甲斐荘,三宅(共著): "新生化学実験講座I,タンパク質III 高次構造" 東京化学同人, 436 (1990)

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      1990 Annual Research Report

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Published: 1990-04-01   Modified: 2016-04-21  

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