Studies of cytochrome c oxidase-driven H(+)-coupled phosphate transport catalyzed by the Saccharomyces cerevisiae Pho84 permease in coreconstituted vesicles.
The proton-coupled Pho84 phosphate permease of Saccharomyces 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.
Fristedt
U
U
Wallenberg Laboratory, Department of Biochemistry, Stockholm University, S-106 91 Stockholm, Sweden.
van Der Rest
M
M
Poolman
B
B
Konings
W N
WN
Persson
B L
BL
D001692:Biological Transport; D002352:Carrier Proteins / Q000096:biosynthesis / Q000378:metabolism*; D002413:Cations, Divalent; D004492:Edetic Acid; D003576:Electron Transport Complex IV / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D008081:Liposomes; D008564:Membrane Potentials; D028044:Phosphate-Binding Proteins; D010710:Phosphates / Q000378:metabolism; D010957:Plasmids; D028081:Proton-Phosphate Symporters; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D029701:Saccharomyces cerevisiae Proteins
D002352:Carrier Proteins; D002413:Cations, Divalent; D008081:Liposomes; C105495:PHO84 protein, S cerevisiae; D028044:Phosphate-Binding Proteins; D010710:Phosphates; D028081:Proton-Phosphate Symporters; D029701:Saccharomyces cerevisiae Proteins; D004492:Edetic Acid; D003576:Electron Transport Complex IV