Simple and rapid purification of F1-ATPase from bovine heart mitochondria by affinity chromatography.
A simple and rapid method for the isolation of bovine heart mitochondrial adenosine 5'-triphosphatase (F1-ATPase) was developed. Mitochondria were purified by differential centrifugation and stored frozen. After thawing. F1-ATPase was released by treatment with chloroform. Purification of the enzyme was achieved by polyethylene glycol precipitation followed by chromatography on Procion Navy H-ER beaded cellulose in the presence of MgCl2. F1-ATPase was eluted by ATP in the absence of MgCl2. The purity of the enzyme was proved by SDS-polyacrylamide-gel electrophoresis. The purified F1-ATPase showed slightly non-hyperbolic kinetics towards ATP and nearly complete inhibition in the presence of millimolar concentrations of ADP.
Temesgen
B
B
Institute of Biochemistry, Karl Marx University, Leipzig, FRG.
Eschrich
K
K
Hofmann
E
E
D000244:Adenosine Diphosphate / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002417:Cattle; D002846:Chromatography, Affinity; D004396:Coloring Agents / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D007700:Kinetics; D015636:Magnesium Chloride / Q000378:metabolism; D008929:Mitochondria, Heart / Q000201:enzymology*; D006180:Proton-Translocating ATPases / Q000302:isolation & purification* / Q000378:metabolism; D014227:Triazines / Q000378:metabolism
D004396:Coloring Agents; D014227:Triazines; D015636:Magnesium Chloride; C003213:Procion Brilliant Red M-2BS; C012842:Cibacron Blue F 3GA; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D006180:Proton-Translocating ATPases