Studies of cytochrome c oxidase-driven H+-coupled phosphate transport catalyzed by the Saccharomyces cerevisiae Pho84 permease in coreconstituted vesicles

被引:27
作者
Fristedt, U
van der Rest, M
Poolman, B
Konings, WN
Persson, BL [1 ]
机构
[1] Stockholm Univ, Dept Biochem, Wallenberg Lab, S-10691 Stockholm, Sweden
[2] Vaxjo Univ, Dept Engn & Nat Sci, S-35195 Vaxjo, Sweden
[3] Univ Groningen, Dept Microbiol, Groningen Biomol Sci & Biotechnol Inst, NL-9751 NH Haren, Netherlands
关键词
D O I
10.1021/bi991545c
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The proton-coupled Pho84 phosphate permease of Saccharomyces cel cerevisiae, overexpressed as a histidine-tagged chimera in Escherichia coli, was detergent-solubilized, purified, and reconstituted into proteoliposomes. Proteoliposomes containing the Pho84 protein were fused with proteoliposomes containing purified cytochrome c oxidase from beef heart mitochondria. Both components of the coreconstituted system were functionally incorporated in tightly sealed membrane vesicles in which the cytochrome c oxidase-generated electrochemical proton gradient could drive phosphate transport via the proton-coupled Pho84 permease. The metal dependency of transport indicates that a metal-phosphate complex is the translocated substrate.
引用
收藏
页码:16010 / 16015
页数:6
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