Structural properties of the proton translocating complex of the clathrin-coated vesicle.
The clathrin-coated vesicle proton pump is a representative member of the new class of endomembrane proton ATPases that share an inhibitor profile which distinguishes them from classic F1F0 and E1E2-type proton pumps. The coated vesicle proton pump is a large (530 kDa) heteroligomer composed of eight polypeptides with molecular masses of 116, 70, 58, 40, 38, 34, 33 and 17 kDa. The 200-fold purified enzyme catalyses ATP-generated proton pumping when reconstituted in liposomes composed of pure lipids. Subunit function has been determined by partial reaction analysis of subunit and subcomplex activities. The isolated 17 kDa subunit, when co-reconstituted with bacteriorhodopsin, forms a dicyclohexylcarbodiimide-inhibitable proton channel. Selective removal of the 116 kDa subunit transforms the proton ATPase from a Mg2+-activatable to a Ca2+-activatable ATPase. Subsequent dissociation and reconstitution of subunits reveals that the 70, 58, 40 and 33 kDa components are required, in composite, to form a functional ATP-hydrolytic core, and that no single subunit or subcomplex deficient in these subunits can catalyse ATP hydrolysis.
Stone
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Department of Physiology, University of Texas Southwestern Medical Center, Dallas 75235-9030.
Sun
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Xie
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D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001436:Bacteriorhodopsins / Q000378:metabolism; D002118:Calcium / Q000494:pharmacology; D002417:Cattle; D002966:Clathrin; D003034:Coated Pits, Cell-Membrane / Q000201:enzymology*; D004024:Dicyclohexylcarbodiimide / Q000494:pharmacology; D011992:Endosomes / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D007473:Ion Channels / Q000187:drug effects / Q000201:enzymology*; D007700:Kinetics; D046911:Macromolecular Substances; D008274:Magnesium / Q000494:pharmacology; D008970:Molecular Weight; D006180:Proton-Translocating ATPases / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism; D011522:Protons
D002966:Clathrin; D007473:Ion Channels; D046911:Macromolecular Substances; D011522:Protons; D001436:Bacteriorhodopsins; D004024:Dicyclohexylcarbodiimide; D000255:Adenosine Triphosphate; D006180:Proton-Translocating ATPases; D008274:Magnesium; D002118:Calcium