1992 年度実績報告書

細菌が生産するプロテアーゼインヒビター機能構造についての蛋白工学的研究

研究課題

研究課題/領域番号	02305003
研究機関	学習院大学
研究代表者	三浦謹一郎学習院大学, 理学部, 教授 (30000227)
研究分担者	赤坂一之神戸大学, 理学部, 教授 (50025368) 外村辨一郎京都大学, 農学部, 教授 (20026545) 甲斐荘正恒東京都立大学, 理学部, 教授 (20137029) 太田隆久東京大学, 農学部, 教授 (30011844) 三井幸雄長岡技術科大学, 工学部, 教授 (40012637)
キーワード	蛋白工学 / 酵素阻害剤 / 阜ロテイーゼ・インヒビター / 部位指定変異 / バクテリア / 遺伝子工学 / 蛋白質の機能部位 / 蛋白質の構造
研究概要	蛋白分解酵素プロテアーゼとそのインヒビター蛋白質の相互作用を分子または分子下レベルで研究した。放線菌のプロテアーゼ・インヒビターSSIは日本で発見され、集中的に研究された蛋白質の一つであるが、この蛋白質を中心にプロテアーゼ・インヒビターを遺伝子操作で改変してプロテアーゼとの相互作用の変化を追究した。SSI遺伝子をクローニングし、作用部位その他の改変を行なった。反応部位の中心を基質と同様なアミノ酸に置換すると、それが接触し易い酵素に対してインヒビターとしての役割を果たすことを明らかにした。また、反応部位以外の部分を改造してプロテアーゼとの接触にどのように効果を挙げることができるかを調べた。さらに2システイン残基によるS-Sの架橋をアミノ酸の置換によってとり払うと、SSIはもはや阻害剤でなくて基質になってしまうことがわかった。野生型ではしっかりした三次元構造をとれるようになっているSSIにそれを破壊するような変化を与えたとき、阻害活性は低下し、インヒビターが基質に転化してしまうことを見いだしたこの結果により、プロテアーゼ・インヒビターの設計のための原理が明らかにされた。SSIの改変体と種々の酵素との複合体の結晶のX線解析からは、プロテアーゼとインヒビターの接触の具合を明らかにした。また、同位元素置換したSSIの構造をNMRで調べると、反応部位付近とC末端寄りの部分がプロテアーゼとの接触において構造変化を起こして易く、誘導適合を施こし得ることを示した。野生型SSIとアミノ酸置換変異体の熱安定性の比較からはSSIの高次構造に大きな変化がないのにもかかわらず、特定位置については協同的なゆうぎが増加していることが示された。この他に放線菌や他の細菌、かび、レトロウィルスあるいは植物種子のプロテアーゼと蛋白性インヒビターの相互作用及び夫々の物性につき多角的に研究を行った。

研究成果
(64件)

すべてその他

すべて文献書誌 (64件)

[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] K.Uchida and M.Kainosho: "Synthesis of [1,1'- ^<13>C_2]-L-Cystine" J.Labelled Compounds and Radiopharmaceuticals. XXIX. 867-874 (1991)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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.
[文献書誌] 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)
[文献書誌] T.Kumazaki and S.Ishii: "Disulfide Bridge Structure of Ascidian Trypsin Inhibitor I:Similarity to Kazal-Type Inhibitors" J.Biochemistry. 107. 414-419 (1990)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] K.Oda and S.Murao: "PEPSTATIN INSENSITIVE CARBOXYL PROTEINASES" Stucture and Function of the Aspartic proteinases. 185-201 (1991)
[文献書誌] 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)
[文献書誌] D.Tsuru,T.Yoshimoto,N.Koriyama and S.Furukawa: "Thiazolidine Derivatives as Potent Inhibitors Specific for Prolyl Endopeptidase" J.Biochemistry. 104. 580-586 (1988)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)
[文献書誌] 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)

1992 年度 実績報告書

細菌が生産するプロテアーゼインヒビター機能構造についての蛋白工学的研究

研究代表者

三浦 謹一郎 学習院大学, 理学部, 教授 (30000227)

研究成果

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] K.Uchida and M.Kainosho: "Synthesis of [1,1'- ^<13>C_2]-L-Cystine" J.Labelled Compounds and Radiopharmaceuticals. XXIX. 867-874 (1991)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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.

[文献書誌] 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)

[文献書誌] T.Kumazaki and S.Ishii: "Disulfide Bridge Structure of Ascidian Trypsin Inhibitor I:Similarity to Kazal-Type Inhibitors" J.Biochemistry. 107. 414-419 (1990)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] K.Oda and S.Murao: "PEPSTATIN INSENSITIVE CARBOXYL PROTEINASES" Stucture and Function of the Aspartic proteinases. 185-201 (1991)

[文献書誌] 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)

[文献書誌] D.Tsuru,T.Yoshimoto,N.Koriyama and S.Furukawa: "Thiazolidine Derivatives as Potent Inhibitors Specific for Prolyl Endopeptidase" J.Biochemistry. 104. 580-586 (1988)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

[文献書誌] 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)

1992 年度実績報告書

三浦謹一郎学習院大学, 理学部, 教授 (30000227)

[文献書誌] 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)