2003 Fiscal Year Final Research Report Summary
Forefront X-Ray Crystallography of Enzymes by using the third generation synchrotron radiation
| Project/Area Number |
13460046
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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 |
応用微生物学・応用生物化学
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| Research Institution | Kyoto University (2002-2003)
The Institute of Physical and Chemical Research (2001) |
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Principal Investigator |
KATO Hiroaki KATO,Hiroaki, 薬学研究科, 教授 (90204487)
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| Co-Investigator(Kenkyū-buntansha) |
HIRATAKE Jun Institute for Chemical Research, Associate Professor, 化学研究所, 助教授 (80199075)
YAMASHITA Atsuko Kyoto University, Structural Biochemistry Laboratory, Researcher, 構造生物化学研究室, 研究員 (10321738)
NAKAMATSU Toru Kyoto University, Graduate School of Pharmaceutical Sciences, Associate Professor, 薬学研究科, 助教授 (50293949)
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| Project Period (FY) |
2001 – 2003
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| Keywords | Time-resolved crystallography / Ultra-high resolution crystallography / Synchrotron radiation / Membrane protein / Laue diffraction / Protein crystallization / Structural biology / Enzyme reaction |
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| Research Abstract |
1.time-resolved x-ray crystallography A combination of two kinetic crystallographic techniques, a continuous flow of the substrates and Laue diffraction measurement, enabled us to capture the transit structures prior to the reaction proceeding. We determined the crystal stuructre of enzyme-substrate complex prior to reaction initiation: tropinone reductase-II-NADPH-tropinone. complex. A structural comparison of the enzyme-substrate complex elucidated in this study with the enzyme-product complex in our previous study indicated that one of the substrates, tropinone, is rotated relative to the product so as to make the spatial organization in the active site favorable for the reaction to proceed.
2.ultra-high-resolution X-ray crystallography Crystal structures of endopolygalacturonase from Stereum purpureum were solved in native and two galacturonic acid complex states at atomic resolution. The active-site architecture of the complexes provides insight into the mechanism of inverting glycosyl hydrolases and also sheds light on the basis of the differences between the family 28 and the other inverting glycosyl hydrolases.
3.X-ray crystal structure determination with difficult analysis We determined crystal structure of maize pyruvate phosphate dikinase, a MARCKS peptide containing the calmodulin-binding domain in complex with Ca(2+)-calmodulin, a myristoylated CAP-23/NAP-22N-terminal domain complexed with Ca(2+)calmodulin, and the PsbP protein of photosystem II from Nicotiana tabacum.
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