| Project/Area Number |
10680639
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Biophysics
|
|---|
| Research Institution | Institute of Technology |
|---|
Principal Investigator |
ITO(SHINZWA) Kyouko Depart. of Life Science, Institute for Protein Research Osaka U., 理学部, 助手 (70206316)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
YAMASHITA Eiki Depart. of Life Science, Institute for Protein Research Osaka U., 蛋白質研究所, 助手 (00294132)
|
|---|
| Project Period (FY) |
1998 – 1999
|
| Project Status |
Completed (Fiscal Year 1999)
|
| Budget Amount *help |
¥3,300,000 (Direct Cost: ¥3,300,000)
Fiscal Year 1999: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1998: ¥2,300,000 (Direct Cost: ¥2,300,000)
|
|---|
| Keywords | Ubiquinol-cyt c reductase / membrane protein / crystallization / Ubiquinone analogue / 呼吸阻害剤 |
|---|
| Research Abstract |
Ubiquinol-cytochrome c reductase (E.C.10.2.2), bc1 complex, is one of three electron transfer complexes which pump proton in the respiratory chain of aerobic organisms. Solving the three dimensional structure of this enzyme at atomic resolution has been the most important subject for a long time for elicidation of the reaction mechanism of this enzyme. Four groups had tried to crystallize this enzyme and recently three goops repotted the crystal structure of the bc1 complex at 2.9A resolution. For elicidation of the reaction mechanism of this enzyme compexes, the crystal structures at high resolution (at least 2.3A resolution) are indispensable.
We have been searching crystallization conditions of the bc1 complexes purified from bovine heart muscle. The detergent structures are important for crystals. We used many type of detergent to stabilize the bc1 complex. Among them, Cychlohexyl-hexyl-maltoside was the most effective for stabilizing the enzyme. Addition of Cychlohexyl-butyl-maltoside to Cychlohesyl-hexyl-maltoside increase the stability of crystals, and the crystals obtained from the samples stabilizing with both detergents diffracted x-ray up to 2.4A resolution.
Stoichiometric excesses of ubiquinome analogue inhibitors, myxothiazol, antimysin, stigmatellin, were added to the enzyme stoichiometric excesses and the crystals bound these inhibitors were obtained. These crystals were diffracted x-ray up to 2.6A resolution. With these crystals, we have obtained the crystallographic data of these native crystals.
|
