1996 Fiscal Year Final Research Report Summary
Structural analysis of the bacterial flagellar switch proteins
| Project/Area Number |
08458207
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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 |
Biophysics
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| Research Institution | Nagoya University |
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Principal Investigator |
OOSAWA Kenji Nagoya University, Graduate School of Polymathematics, associate professor, 大学院・多元数理科学研究科, 助教授 (50203758)
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| Project Period (FY) |
1996
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| Keywords | bacteria / flagellum / flagellar motor / switch protein / structural analysis / protein crystal / overproduction / molecular machine |
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| Research Abstract |
For structural analysis of the bacterial flagellar switch proteins, we examined conditions for purification and crystallization of the proteins. The FliN protein was highly purified by the published procedures. Therefore, we have been trying to find conditions for crystallization of the purified protein. However, from this sample we sometimes could not get any spherolyte. Because we do not have any method to check whether the protein is intact, it was not known whether the purified protein was homogenous. We have started using the new procedure for purification. In this procedure, the overproduced FliN was in soluble fraction in the cells, whereas in the former case the protein in insoluble form. By the new method, we can purify the FliN protein which forms spherolytes. On the other hand, for the FliM protein, it is very difficult to solubilize the protein. We are now planning construction of overproducing plasmids for producing fragments of the FliM containing functional domains. These FliM fragments may possibly be better for solubilization and crystallization. Also, we have been working to determine overproducing conditions for the FliM and FliN proteins, e. g. temperature, pH,etc., for solubilizing these proteins. Because in some cases overproducing proteins was shown not to form inclusion bodies in certain overproducing conditions.
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