Microheterogeneity of rat glycogen phosphorylase liver-type isozyme

Abstract

We devised a method of polyacrylamide gel electrophoresis at pH 7.3, modified by omitting base catalyst, N,N,N′,N′- tetramethylethylenediamine , in the preparation of separating gels. Using this method, both liver and liver-like types of rat glycogen phosphorylase (1,4-α- d-glucan:orthophosphate α-glucosyltransferase, EC 2.4.1.1) were resolved into multiple forms, about 6–10, although either of them was purified to a single protein with the same molecular size on sodium dodecyl sulfate gel electrophoresis. The microheterogeneity of these two types was also confirmed by isoelectric focusing in polyacrylamide gels (pH 5–8). The major isoelectric points of the liver type phosphorylase were between 5.72 and 5.86, but those of the liver-like type were between 5.86 and 5.92, and so the former had slightly but significantly lower isoelectric points that the latter. However, the both types were not distinguished immunologically. The brain and muscle types of rat phosphorylase did not show such a distinct heterogeneity by the same electrophoresis methods.