Reactivation of proteins and membrane transport by the heat shock proteins from thermophilic bacteria.

嗜热细菌的热休克蛋白重新激活蛋白质和膜运输。

• 批准号: 01870015
• 负责人: KAGAWA Yasuo
• 金额: $ 19.26万
• 依托单位: Jichi Medical School
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research
• 财政年份: 1989
• 资助国家: 日本
• 起止时间: 1989 至 1991
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-01870015/
• 关键词: heat shock protein; thermophilic chaperonin; reconstitution; GroEL; thermophilic gene; hsp gene; ATP synthase; oligomer; synchrotron radiation; chaperonin; biomembrane; protein renaturation; conformationーbuildーup; membrane transport; thermop

Heat shock proteins are essential for tlie formation and restoration of many supramolecular structures. For this purpose, we isolated stable heat shock proteins of 6lkDa, 12kDa and 69kDa, which are similar to GroEL, GroES and DnaK, respectively, 'from the supernatant fraction of thermophilic bacterium PS3 by ATP-Agarose chromatography. Using synthetic DNA of the deduced sequence, the 1.6kbp DNA encoding both proteins was obtained by polymerase chain reaction. The complete sequence of the resulting reading frames showed high. homology to those of the genes encoding GroEL and GroES, and the 12kDa and 6lkDa genes were present in the same operon. 61KDa was also partially similar to the F_1alpha subunit of thermophilic ATP synthase. Using the over-expressed thermophilic F_1alpha subunit it was possible to isolate the alpha_3beta_3 and alpha_1beta_1 subunit complexes. These results subunits have been analyzed by synchrotron radiation etc. These results are precious to analyze energy transformation in the reconstituted membrane system.Patent : Toku-Gan-Hei#2-73664, Heat shock protein, Applicant : Yasuo Kagawa.

热休克蛋白是许多超分子结构的形成和恢复所必需的。为此，我们利用atp -琼脂糖层析法从嗜热细菌PS3的上清部分分离出与GroEL、GroES和DnaK相似的6lkDa、12kDa和69kDa稳定的热休克蛋白。利用推导出的序列合成DNA，通过聚合酶链反应获得编码这两个蛋白的1.6kbp DNA。所得到的阅读帧的完整序列显示为高。与编码GroEL和GroES的基因同源，12kDa和6lkDa基因存在于同一操纵子中。61KDa也部分类似于嗜热ATP合成酶的f_1 α亚基。利用过表达的嗜热性f_1 α亚基，可以分离出alpha_3beta_3和alpha_1beta_1亚基复合物。用同步辐射等方法对这些结果进行了分析。这些结果对分析重组膜系统的能量转化具有重要的参考价值。专利号：Toku-Gan-Hei#2-73664，热休克蛋白，申请人：香川康夫。

项目成果

Tamada, H., Ohta, T., Hamamoto, T., Otawara - Hamamoto, Y., Yanagi, M., Hiraiwa, H., Hirata, H. and Kagawaw, Y.: "Gene structure of heat shock proteins 61KDa and 12KDa (thermophilic chaperonins) of thermophilic bacterium PS3." Biochemical and Biophysical

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