Proton translocating ATPase of a thermophilic bacterium. Morphology, subunits, and chemical composition.
1. A stable membrane-bound ATPase [EC 3.6.1.3] (TF0-F1) capable of proton translocation in reconstituted vesicles was purified from the thermophilic bacterium PS3 cultured in medium containing L-[U-14C]amino acids. 2. TF0-F1 was composed of a catalytic moiety (TF1) and a hydrophobic moiety (TF0). TF1 contained 3 polypeptide chains with molecular weights of 56,000, 3 of 53,000, 1 of 32,000, 1 of 15,500, and 1 of 11,000. TF0 contained 1 chain of 19,000, 2 of 13,500, and 5 of 5,400 daltons. TF1 was dissociated into subunits much less readily than F1. 3. TF1 consisted of 95A particles arrayed in hexagonal microcrystals. TF0-F1 consisted of a sphere (TF1) and a stalk plus base (TF0) which was buried in the membrane of the proton translocating vesicles. 4. Vesicles capable of energy transformation were formed when TF1 came in contact with the surface of liposomes containing TF0. On addition of phospholipids, the helix content of TF0 increased 3-fold. The role of F0 in forming channels for protons is discussed. 5. The amino acid compositions of TF0, TF1, and TF0-F1 were compared. TF0 was not hydrophobic, despite its interaction with phospholipids. The phospholipid composition and other properties of the proton translocating vesicles were examined. Vesicles reconstituted from a mixture of phosphatidylethanolamine, phosphatidylgly-cerol, and cardiolipin in the same ratio as in the membranes had the highest activity.
D000251:Adenosine Triphosphatases / Q000378:metabolism*; D000596:Amino Acids / Q000032:analysis / Q000378:metabolism; D001419:Bacteria / Q000201:enzymology*; D002462:Cell Membrane / Q000201:enzymology; D004024:Dicyclohexylcarbodiimide / Q000494:pharmacology; D004735:Energy Transfer; D005227:Fatty Acids / Q000032:analysis; D008094:Lithium / Q000494:pharmacology; D008970:Molecular Weight; D010446:Peptide Fragments / Q000032:analysis; D010743:Phospholipids / Q000494:pharmacology; D011487:Protein Conformation
D000596:Amino Acids; D005227:Fatty Acids; D010446:Peptide Fragments; D010743:Phospholipids; D004024:Dicyclohexylcarbodiimide; D008094:Lithium; D000251:Adenosine Triphosphatases