分裂泛素化酵母双杂交系统研究光合膜蛋白相互作用  

Studying of The Thylakoid Membrane Protein Interactions Through The Split-ubiquitin Yeast Two Hybrid System

作　　者：朱树勇[1] 迟伟[1] 张立新[1] 

机构地区：[1]中国科学院植物研究所光合作用与环境分子生理重点实验室

出　　处：《生物化学与生物物理进展》2006年第4期350-356,共7页Progress In Biochemistry and Biophysics

基　　金：国家自然科学基金资助项目(30500037);中国科学院植物研究所特别支持经费.

摘　　要：光合类囊体膜主要由光系统Ⅱ、细胞色素b6f复合物、光系统Ⅰ以及ATP合酶4个超分子复合物组成.利用分裂泛素化酵母双杂交系统研究光合类囊体膜蛋白间的相互作用.将叶绿体psbA基因编码的D1蛋白作为诱饵蛋白,以叶绿体基因psbD编码的D2蛋白、petB编码的Cytb6蛋白作为靶蛋白,分别共转化酵母菌株后进行相互作用分析.实验结果表明,诱饵蛋白D1能与来源于同一复合物光系统Ⅱ的D2蛋白发生相互作用,而与来源于细胞色素b6f复合物的Cytb6蛋白没有互作.这一结果表明,分裂泛素化酵母双杂交系统可以用于检测光合膜蛋白间的相互作用,从而为研究光合膜蛋白生物发生的调控机理提供一个有效的工具.The major photosynthetic complexes PS Ⅰ, PS Ⅱ, the cytochrome b6f complex and the ATP synthase complex are located in the thylakoid membrane. Here, the newly developed split-ubiquitin two hybrid system was used to investigate protein-protein interactions of thylakoid membrane proteins. The reaction center D 1 protein of PS Ⅱ, which is encoded by plastid gene psbA, was used as the bait protein, while the D2 encoded by plastid gene psbD, another PS Ⅱ reaction center protein, and Cytb6 encoded by plastid gene petB, a component of Cytb6f complex, were used as prey proteins. The yeast two hybrid analysis showed that the D1 protein interacts with D2, but not with the Cytb6. This result confirms the protein interaction model of thylakoid membrane protein complex, and also means the availability of this system in detecting thylakoid membrane protein interactions. Thus, the split-ubiquitin two hybrid systems could provide an efficient tool to reveal the regulation mechanism of chloroplast proteins biogenesis.

关 键 词：光合膜蛋白 蛋白质相互作用 分裂泛素化酵母双杂交系统 光系统Ⅱ 

分 类 号：Q71[生物学—分子生物学]