Acetyl-coenzyme-A carboxylase from rat liver. Subunit structure and proteolytic modification.
The subunit structure of rat liver acetyl-coenzyme-A carboxylase has been studied by polyacrylamide gel electrophoresis in the presence of dodecylsulfate. A number of individual preparations of the enzyme purified by the same procedures exhibited three different types of electrophoretic patterns as follows: first, a single slow-moving protein bands (Mr 230000); secondly, two adjacent fast-moving protein band (M4 124000 and 118 000); finally, all three protein bands. With the use of the [14C]biotin-labelled enzyme, the biotinyl prosthetic group was shown to be associated with the polypeptide of 230000 Mr as well as with that of 124000 Mr, but not with the polypeptide of 118000 Mr. Studies were next made with the labelled enzyme to examine the possibility that the two light polypeptides might have been formed by proteolytic modification of the heavy polypeptide during the procedures used for the purification of the enzyme. Treatment of the enzyme with trypsin or chymotrypsin resulted in cleavage of the heavy polypeptide into two nonidentical polypeptides with molecular weights of approximately 120000. Incubation of the enzyme with proteases derived from rat liver converted the heavy polypeptide into lighter polypeptides of 80000-130000 Mr. Acetyl-CoA carboxylase isolated from crude rat liver extracts by means of immunoprecipitation with specific antibody invariably showed only the heavy polypeptide. The biotin content of the enzyme was found to be 1 mol per 237000 g protein. These results indicate that rat liver acetyl-CoA carboxylase, unlike bacterial and plant biotin enzymes, has only one kind of subunit, which has a molecular weight of 230000 and contains one molecular of biotin. Thus, the mammalian enzyme exhibits a highly integrated subunit structure.
D000103:Acetyl-CoA Carboxylase / Q000276:immunology / Q000378:metabolism; D000818:Animals; D001710:Biotin / Q000032:analysis; D004041:Dietary Fats; D004591:Electrophoresis, Polyacrylamide Gel; D008025:Ligases; D008099:Liver / Q000201:enzymology*; D008297:Male; D008970:Molecular Weight; D010455:Peptides / Q000032:analysis; D051381:Rats; D014357:Trypsin
D004041:Dietary Fats; D010455:Peptides; D001710:Biotin; D014357:Trypsin; D008025:Ligases; D000103:Acetyl-CoA Carboxylase