| Project/Area Number |
18380068
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Applied biochemistry
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| Research Institution | Kyoto Institute of Technology |
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Principal Investigator |
HIRAGA Kazumi (2007) Kyoto Institute of Technology, Graduate School of Science and Technology, Assistant Professor (50252549)
小田 耕平 (2006) 京都工芸繊維大学, 工芸科学研究科, 教授 (50081584)
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| Co-Investigator(Kenkyū-buntansha) |
平賀 和三 京都工芸繊維大学, 工芸科学研究科, 助手 (50252549)
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| Project Period (FY) |
2006 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥14,650,000 (Direct Cost: ¥13,600,000、Indirect Cost: ¥1,050,000)
Fiscal Year 2007: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000)
Fiscal Year 2006: ¥10,100,000 (Direct Cost: ¥10,100,000)
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| Keywords | sedolisin / e olisin / elutamic nentidase / serine-carboxvl proteinase / structure and function / substrate specificity / reaction mechanism / proteinase inhibitor / serine-carboxy1 peptidase / プロテアーゼ / ペプチダーゼ / セリン-カルボキシルプロテアーゼ / グルタミン酸プロテアーゼ |
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| Research Abstract |
Sedolisin : 1) Structural analysis of CLN2 subsite : Combinatorial substrate library was designed and synthesized, and subsite-analysis is underway. 2) structure analysis of CLN2 : Procedures for expression and purification were investigated and an extremely efficient purification method was constructed. Large amount of CLN2 was prepared and the crystallization of CLN2 is undergoing.
Eqolisin family: 1) Elucidation of reaction mechanism: Glu136 and G1n53, presumed catalytic residues by structural analysis, were changed to Ala and their reaction kinetics revealed that these residues were the catalytic resideues. Based on the detailed analysis for substrate specificity using combinatorial library, synthetic inhibitor was developed (Y. Kataoka et al: FEBSE Letters, 579, 2991-2994 (2005)). Furthermore, enzyme-inhibitor complex was prepared and the obtained results from the structural analysis revealed the reaction mechanism (B. Pillai, et. al.: J. Mol Biol 365, 343-361 (2007)). 2) Distribution in nature: not completed. 3) Development of safety agrichemicals: Peptide inhibitors having inhibitory constant at subnano-molar-level were successively developed. The effect of the inhibitor for phytopathogenic microbials will be determined at lab scale.
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