Insights into the subunit in-teractions of the chloroplast ATP synthase  

作　　者：Shi, XB Wei, JM Shen, YG 

机构地区：[1]Chinese Acad Sci, Shanghai Inst Biol Sci, Shanghai Inst Plant Physiol, Shanghai 200032, Peoples R China

出　　处：《Chinese Science Bulletin》2002年第1期58-62,共5页

基　　金：This work was supported by theState Key Basic Research and Development Plan (Grant No. G1998010100) ; the National Natural Science Foundation of China (Grant No. 39730040).

摘　　要：Subunit interactions of the chloroplast F0F1-ATP synthase were studied using the yeast two-hybrid sys-tem. The coding sequences of all the nine subunits of spinach chloroplast ATP synthase were cloned in two-hybrid vectors. The vectors were transformed into the yeast strains HF7c and SFY526 by various pairwise combinations, and the protein interactions were analyzed by measuring the yeast growth on minimal SD medium without serine, lucine and histidine. Interactions of γ subunit with wild type or two truncated mutants of ε sununit, ε△N21 and ε△C45, which lose their abilities to inhibit the ATP hydrolysis, were also detected by in vitro and in vivo binding assay. The present results are largely accordant to the common structure model of F0F1-ATP synthase. Different from that in the E. coli F0F1-ATP synthase, the δ subunit of chloroplast ATP synthase could interact with β. γ, ε and all the CF0 subunits in the two-hybrid system. These results suggested that though the chloroplast ATP synthaseSubunit interactions of the chloroplast F0F1-ATP synthase were studied using the yeast two-hybrid sys-tem. The coding sequences of all the nine subunits of spinach chloroplast ATP synthase were cloned in two-hybrid vectors. The vectors were transformed into the yeast strains HF7c and SFY526 by various pairwise combinations, and the protein interactions were analyzed by measuring the yeast growth on minimal SD medium without serine, lucine and histidine. Interactions of γ subunit with wild type or two truncated mutants of ε sununit, ε△N21 and ε△C45, which lose their abilities to inhibit the ATP hydrolysis, were also detected by in vitro and in vivo binding assay. The present results are largely accordant to the common structure model of F0F1-ATP synthase. Different from that in the E. coli F0F1-ATP synthase, the δ subunit of chloroplast ATP synthase could interact with β. γ, ε and all the CF0 subunits in the two-hybrid system. These results suggested that though the chloroplast ATP synthase shares the similar structure and composition of subunits with the enzyme from E. coli, it may be different in the subunit interactions and con-formational change during catalysis between these two sources of ATP synthase. Based on the present results and our knowledge of structure model of E. coli ATP synthase, a deduced structure model of chloroplast ATP synthase was proposed.

关 键 词：F0F1-ATP SYNTHASE SUBUNIT interaction YEAST TWO-HYBRID system δ subunit. 

分 类 号：Q946[生物学—植物学]