Purification and properties of Mg2+-Ca2+ adenosinetriphosphatase from Escherichia coli.
A procedure for the purification of Mg(2+)-Ca(2+) adenosinetriphosphatase (EC 3.6.1.3) from E. coli, yielding relatively large amounts of highly active enzyme, is described. The enzyme consists of four nonidentical subunits. Trypsin treatment of purified enzyme yields a preparation consisting exclusively of the two larger subunits, which are sufficient for ATPase activity. Purified enzyme is inhibited by 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole; this inhibition is reversed by dithiothreitol, and the diazole is found preferentially associated with the beta-subunit of the enzyme. Antibody prepared against the trypsin-treated enzyme inhibited various ATP-dependent reactions as well as membrane-bound ATPase itself.
D000244:Adenosine Diphosphate; D000251:Adenosine Triphosphatases / Q000032:analysis / Q000037:antagonists & inhibitors / Q000302:isolation & purification*; D000255:Adenosine Triphosphate; D000906:Antibodies; D002118:Calcium; D004229:Dithiothreitol; D004586:Electrophoresis; D004926:Escherichia coli / Q000201:enzymology*; D008274:Magnesium; D011485:Protein Binding; D012967:Sodium Dodecyl Sulfate; D013439:Sulfhydryl Reagents; D014316:Tritium; D014357:Trypsin
D000906:Antibodies; D013439:Sulfhydryl Reagents; D014316:Tritium; D012967:Sodium Dodecyl Sulfate; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D014357:Trypsin; D000251:Adenosine Triphosphatases; D008274:Magnesium; D002118:Calcium; D004229:Dithiothreitol