Proton translocation by ATPase and bacteriorhodopsin.
Stable membrane proteins and lipids are convenient to study biomembranes. Two stable proton translocating proteins were purified and reconstituted into vesicles capable of proton translocation. One was a thermostable ATPase (TF0-F1) of thermophilic bacterium PS3 and the other was rhodopsin of Halobacterium halobium. TF0-F1 was composed of a proton pump moiety (TF1) and a proton channel moiety (TF0). TF1 was the first membrane ATPase which was crystallized and reconstituted from its five polypeptides. Like TF0 and TF1, the rhodopsin in purple membrane was highly stable against dissociating agents, acids and alkali. Phospholipids of these biomembranes were also stable and contained no unsaturated fatty acyl groups. The molecular species of the phospholipids of PS3 were determined by mass chromatography. Measurements were made of the difference in electrochemical potential of protons (deltamicronH+) across the membrane of the reconstituted vesicles. The deltamicronH+ attained was 312 mV in TF0-F1 vesciles and was 230 mV in the rhodopsin vesicles. To conclude that electron transport components are not necessary for ATP synthesis in energy yielding biomembranes, two experiments were performed: The ATP synthesis was observed i) on acid-base treatment of TF0-F1 vesicles, and ii) on illumination of the rhodopsin-TF0-F1 vesicles.
D000251:Adenosine Triphosphatases / Q000378:metabolism*; D001436:Bacteriorhodopsins / Q000378:metabolism; D002259:Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone / Q000494:pharmacology; D002338:Carotenoids / Q000378:metabolism; D004024:Dicyclohexylcarbodiimide / Q000494:pharmacology; D006217:Halobacterium / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D006863:Hydrogen-Ion Concentration; D008081:Liposomes; D008854:Microscopy, Electron; D010743:Phospholipids; D011485:Protein Binding; D011487:Protein Conformation; D011510:Proteolipids / Q000032:analysis; D013816:Thermodynamics
D008081:Liposomes; D010743:Phospholipids; D011510:Proteolipids; D002338:Carotenoids; D002259:Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone; D001436:Bacteriorhodopsins; D004024:Dicyclohexylcarbodiimide; D000251:Adenosine Triphosphatases