Research on the Mechanisms of Intra-cellular Transportation and Functional Regulation of GTP Binding Proteins Using Schizosaccharomyces pombe

利用粟酒裂殖酵母研究 GTP 结合蛋白的细胞内运输机制和功能调控

• 批准号: 04833027
• 负责人: FUJIYAMA Asao
• 金额: $ 1.28万
• 依托单位: National Institute of Genetics
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1993
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-04833027/
• 关键词: GTP BINDING PROTEIN; INTRA-CELLULAR LOCALIZATION; POSTTRANSLATIONAL MODIFICATION; FISSION YEAST; ISOPRENYLATION; FARNESYLATION; RAS PROTEIN; ファルネル化; 速度論的解析; ras蛋白質; in vitro再構成; G蛋白質

The purpose of the research is to analyze mechanisms of intracellular-transportation and localization of small molecular weight GTP binding proteins, Fro this purpose, we have been using fission yeast, Schizosaccharomyces pombe, as a model system. In the first year of the research, we purified pombe rasl protein. The biochemical analyzes of the protein revealed that the protein, unlike other ras proteins, has larger Kd value to GDP than to GTP.This is the first report of the biochemical analyzes of pombe rasl protein. We also isolated a new gene that encoded for novel G-protein coupled receptor protein. Knowing the characteristics of the rasl protein, we moved to reconstruct posttranslational modification system in vitro in the second, and it is the last year of the project. From the kinetic analyzes of the partially purified farnesyl transferase, we found out the difference in the rate constant rather than Km value against various peptide substrates. This will be further investigated to clarify the mechanism of substrate selection by various preny1-transferases in vivo.

本研究的目的是分析小分子GTP结合蛋白的胞内转运和定位机制，为此，我们一直使用裂解酵母作为模型系统。在研究的第一年，我们纯化了Pombe RASL蛋白。对该蛋白的生化分析表明，与其他ras蛋白不同，该蛋白对GDP的Kd值大于对GTP的Kd值，这是首次报道了Pombe RASL蛋白的生化分析。我们还分离到了一个编码新的G蛋白偶联受体蛋白的新基因。了解到RASL蛋白的特性，我们在第二个阶段进行了体外翻译后修饰系统的重建，这也是该项目的最后一年。从对部分纯化的法尼基转移酶的动力学分析中，我们发现对不同的多肽底物的速率常数而不是Km值的差异。这将被进一步研究，以阐明体内各种戊烯1-转移酶选择底物的机制。

项目成果

Danjo ら: "The Post translationul modification of Cterminal Amiu Acids" Amino Acids. 5. 147-147 (1993)

Danjo 等人：“C 末端氨基酸的翻译后修饰”《氨基酸》5. 147-147 (1993)。

檀上 ら: "Choroideremia遺伝子はGeranyl geranyl transferaseか" 実験医学. 11. 68-70 (1993)

Danjo 等人：“无脉络膜基因是香叶基香叶基转移酶吗？”实验医学。11. 68-70 (1993)

〓上 ら: "Choroideremia 遺伝子はGerany(gerayltras ferase か" 実験医学. 11. 68-70 (1993)

Kami 等人：“无脉络膜症基因 Gerany(geryltras ferase) 是吗？”《实验医学》11. 68-70 (1993)。

Qadota, HT., Ishii, I., Fujiyama, A., Ohya, Y.and Anraku, Y.: "RHO Gene Products, Putative Small GTP-binding Proteins, are Important for Activation of the CAL1/CDC43 Gene Product, a Protein Geranylgeranytransferase in Saccharomyces cerevisiae." Yeast. 8.

Qadota, HT.、Ishii, I.、Fujiyama, A.、Ohya, Y. 和 Anraku, Y.：“RHO 基因产物，推定的小 GTP 结合蛋白，对于 CAL1/CDC43 基因产物的激活非常重要，

Danjo, I., Iha, H., Onozawa, T., Zhaojun, X.and Fujiyama, A.: "The posttranslational modification of C-termional amino acids of the ras super family proteins." Amino Acids. 5. 147 (1993)

Danjo, I.、Iha, H.、Onozawa, T.、Zhaojun, X. 和 Fujiyama, A.：“ras 超家族蛋白 C 端氨基酸的翻译后修饰。”