A new method for the isolation of rat liver acetyl-CoA carboxylase.
Rat liver acetyl-CoA carboxylase has been purified to homogeneity by a new method involving polyethylene glycol precipitation, and DEAE and Sepharose 4B chromatography. The final product displays a single band on SDS polyacrylamide gel electrophoresis of estimated molecular weight 240,000. This material contains 5.5 +/- 0.3 moles of alkali-labile phosphate per subunit and has a specific activity of 1.2 +/- 0.2 units per mg protein. As compared to previous purification procedures for the liver enzyme, this product has a higher phosphate content, lower specific activity, and an absence of major proteolysis. Trypsin digestion of 32P-labeled acetyl-CoA carboxylase from hepatocytes reveals that the 32P-labeled phosphorylation sites are extremely labile to proteolytic digestion. Potential modification of isolated liver acetyl-CoA carboxylase by proteolysis and/or dephosphorylation must be ascertained prior to in vitro enzymatic studies.
D000103:Acetyl-CoA Carboxylase / Q000302:isolation & purification*; D000818:Animals; D055598:Chemical Phenomena; D011232:Chemical Precipitation; D002621:Chemistry; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D008025:Ligases / Q000302:isolation & purification*; D008099:Liver / Q000201:enzymology*; D008297:Male; D008722:Methods; D008970:Molecular Weight; D011092:Polyethylene Glycols; D011817:Rabbits; D051381:Rats; D014357:Trypsin
D011092:Polyethylene Glycols; D014357:Trypsin; D008025:Ligases; D000103:Acetyl-CoA Carboxylase