Purification of a soluble ATPase from rat liver mitochondria by AMP-Sepharose affinity chromatography.
ATPase (ATP phosphohydrolase, EC 3.6.1.3) activity was shown in the soluble fraction of rat liver micochondria. Two molecular forms (ATPase 1 and 2) were isolated. ATPase 1 has already been studied. The present paper deals with the purification method of ATPase 2 which was achieved by the following steps: (NH4)2SO4 precipitation. DEAE-cellulose chromatography, hydroxyapatite chromatography, Sephadex G100 filtration and AMP-Sepharose affinity chromatography. The purified protein was characterized by bidimensional polyacrylamide gel electrophoresis. Molecular weight evaluated by SDS-polyacrylamide gel electrophoresis and Sephadex G100 gel filtration was found to be 61 500 +/- 3000.
D000249:Adenosine Monophosphate; D000251:Adenosine Triphosphatases / Q000302:isolation & purification*; D000818:Animals; D002846:Chromatography, Affinity; D008722:Methods; D008930:Mitochondria, Liver / Q000201:enzymology*; D008970:Molecular Weight; D051381:Rats; D012685:Sepharose
D000249:Adenosine Monophosphate; D012685:Sepharose; D000251:Adenosine Triphosphatases