A NEW METHOD FOR THE ISOLATION OF RAT-LIVER ACETYL-COA CARBOXYLASE
- 1 January 1981
- journal article
- research article
- Vol. 22 (2), 364-369
Abstract
Rat liver acetyl-CoA carboxylase was 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 [sodium dodecylsulfate] polyacrylamide gel electrophoresis of estimated MW 240,000. This material contains 5.5 .+-. 0.3 mol of alkali-labile phosphate/subunit and has a specific activity of 1.2 .+-. 0.2 U/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.This publication has 10 references indexed in Scilit:
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