The ATP synthetase of Escherichia coli K12: purification of the enzyme and reconstitution of energy-transducing activities.
The ATP synthetase of Escherichia coli K12 was purified by a simple procedure. The dicyclohexylcarbodiimide-sensitive ATPase activity was enriched 21-fold. The ATP synthetase preparation contained the eight polypeptides (alpha, beta, gamma, a,delta, b,espilon, c) of the enzyme and a residual contamination (4% of the total protein) as shown by dodecylsulfate/polyacrylamide electrophoresis. The polypeptide c was specifically labelled with [14C]dicyclohexylcarbodiimide. Energy-transducing activities were reconstituted from soybean phospholipids and the purified enzyme. The proteoliposomes exhibited a significantly higher ATP-32Pi exchange activity and a higher proton-translocating activity as compared to the untreated membranes.
D000244:Adenosine Diphosphate; D004024:Dicyclohexylcarbodiimide; D004735:Energy Transfer; D004926:Escherichia coli / Q000201:enzymology*; D007700:Kinetics; D046911:Macromolecular Substances; D009097:Multienzyme Complexes / Q000302:isolation & purification / Q000378:metabolism*; D010770:Phosphotransferases / Q000302:isolation & purification / Q000378:metabolism*; D011485:Protein Binding
D046911:Macromolecular Substances; D009097:Multienzyme Complexes; D004024:Dicyclohexylcarbodiimide; D000244:Adenosine Diphosphate; D010770:Phosphotransferases