1999 Fiscal Year Final Research Report Summary
Development of high pressure cells for circular dichroism and high pressure effects on protein secondary structure.
| Project/Area Number |
10558097
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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 | 展開研究 |
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| Research Field |
Structural biochemistry
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| Research Institution | KYOTO UNIVERSITY |
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Principal Investigator |
HAYASHI Rikimaru Kyoto University, Professor, 農学研究科, 教授 (90027186)
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| Co-Investigator(Kenkyū-buntansha) |
KAWAI Shoji Teramecs Company Ltd., 開発部, 主任研究員
SATOU Shuuichi SVMITOMO ELECTRIC INDUSTRIES, LTD, 研究開発本部, 主任
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| Project Period (FY) |
1998 – 1999
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| Keywords | high pressure / circular dichroism / carboxypeptidase Y / ribonuclease A / diamond windows / pressure tolerance / high pressure circular dichroism / high pressure spectroscopy |
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| Research Abstract |
To establish in situ observation of high pressure spectroscopy in the field of bioscience and biotechnology, convenient and routine cells for circular dichroism (CD) were developed and pressure dependent changes of α, β and random structures in poly-glutamic acid and poly-lysine were studied. These studies were extended to measure the pressure effects on ribonuclease A and carboxypeptidase Y and their genetically mutated forms. The results are as follows:
1. Pressure tolerance and absorbance of ultra violet light were tested with quartz, sapphires, and artificial diamonds as windows of high pressure CD cells. As the results, the diamond window was found to be applicable to measure CD from 230 to visible light region.
2. Pressure effects on α, β and random structures of poly-glutamic acid and poly-lysine were measured under various pH. Unfortunately, since diamond windows absorbed strongly ultra violet light at 220 nm, clear description of spectrum was difficult.
3. Pressure effects on bovine pancreatic ribonuclease A and carboxypeptidase Y were measured under various pH and temperature and their Pm were determined. Pm was also determined with mutant enzymes.
4. Pressure effects of CD at 280 nm were measured with ribonuclease A and the pressure dependent change of the tertiary structure was proved to be the two state transition as the temperature dependent change.
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