| Project/Area Number |
14560060
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
応用微生物学・応用生物化学
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| Research Institution | Shinshu University |
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Principal Investigator |
SHIDA Toshio Shinshu University, Faculty of Textile Science and Technology, Associate Professor, 繊維学部, 助教授 (40162599)
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| Co-Investigator(Kenkyū-buntansha) |
SEKIGUCHI Junichi Shinshu University, Faculty of Textile Science and Technology, Professor, 繊維学部, 教授 (80111053)
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| Project Period (FY) |
2002 – 2003
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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 2003: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 2002: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Keywords | Bacillus subtilis / cell wall / peptideglycan / amidase / binding domain / NMR structure / 立体構造 / 細胞壁溶解酵素 |
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| Research Abstract |
The Bacillus subtilis CwlC is the cell wall lytic N-acetylmuramoyl-l-alanine amidases in the CwlB (LytC) family that contains a homologous catalytic domain. The enzymes are thought to play an important role in mother-cell lysis in sporulation. The CwlC consists of a N-terminal catalytic domain and a tandem repeat (repeat-1 : 184-219 and repeat-2 : 220-254) in the C-terminal region. Biochemical analysis has shown that the C-terminal tandem repeat, named as CwlCr, can bind to the B. subtilis peptidoglycan (unpublished).
We tried to determine the structure of CwlCr for understanding a peptidoglycan binding mechanism. Using standard multi-dimensional hetero nuclear NMR methods, we completed the main-chain and side-chain resonance assignments, and collected distance restraints and dihedral angle restraints. Furthermore, unambiguous 26 hydrogen bond restraints were obtained from HNCO(^<h3>J_<NC>) experiment. Structure calculation was performed using CYANA, and a low resolution structure was obtained so far. Intriguingly, the each repeat adopted β α β structure making a β-sheet between repeat1 and repeat2. Thus, it was likely that both repeat-1 and repeat-2 were required for CwlCr folding. The refinement process of structure calculation and mutation analyses are under way. We will discuss the interaction of CwlCr with peptidoglycan in detail based on an NMR titration experiment.
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