Phosphorylation of rat liver glycogen synthase bound to the glycogen particle.
Rat liver glycogen synthase bound to the glycogen particle was partially purified by repeated high-speed centrifugation. This synthase preparation was labeled with 32P by incubations with cAMP-dependent protein kinase and cAMP-independent synthase (casein) kinase-1 in the presence of [gamma-32P]ATP. The phosphorylated synthase was separated from other proteins in the glycogen pellet by immunoprecipitation with rabbit anti-rat liver glycogen synthase serum. Analysis of the immunoprecipitates by sodium dodecyl sulfate-gel electrophoresis showed that synthase subunits of Mr 85,000 and 80,000 were present in varying proportions. The 32P-labeled synthase in the immunoprecipitate was digested with trypsin, and the resulting peptides were analyzed by isoelectric focusing. Synthase bound to the glycogen particle was phosphorylated by cAMP-dependent protein kinase at more sites and by cAMP-independent synthase (casein) kinase-1 at less sites than when the homogeneous synthase was incubated with these kinases. Phosphorylation of synthase in the glycogen pellet by either cAMP-dependent protein kinase or cAMP-independent synthase (casein) kinase-1 did not cause a significant inactivation as has been observed when the synthase was incubated with these kinases. Inactivation of synthase in the glycogen pellet, however, can be achieved by the combination of both kinases. This inactivation appears to result from the phosphorylation of a new site by cAMP-independent synthase (casein) kinase-1 neighboring a site previously phosphorylated by cAMP-dependent protein kinase.
D000818:Animals; D000242:Cyclic AMP / Q000378:metabolism; D004533:Egtazic Acid / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D006003:Glycogen / Q000378:metabolism*; D006006:Glycogen Synthase / Q000378:metabolism*; D006657:Histones / Q000494:pharmacology; D007525:Isoelectric Focusing; D008099:Liver / Q000201:enzymology*; D008970:Molecular Weight; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism; D051381:Rats; D014357:Trypsin / Q000378:metabolism
D006657:Histones; D004533:Egtazic Acid; D006003:Glycogen; D000242:Cyclic AMP; D006006:Glycogen Synthase; D011494:Protein Kinases; D014357:Trypsin