title: Phosphorylation of sites 3 and 2 in rabbit skeletal muscle glycogen synthase by a multifunctional protein kinase (ATP-citrate lyase kinase).
abstract: A multifunctional protein kinase, purified from rat liver as ATP-citrate lyase kinase, has been identified as a glycogen synthase kinase. This kinase catalyzed incorporation of up to 1.5 mol of 32PO4/mol of synthase subunit associated with a decrease in the glycogen synthase activity ratio from 0.85 to a value of 0.15. Approximately 65-70% of the 32PO4 was incorporated into site 3 and 30-35% into site 2 as determined by reverse phase high performance liquid chromatography. Release of 32PO4 from the phosphopeptides during automated Edman degradation confirmed the site 3 and 2 assignment. Thermal stability studies established that the phosphorylations of sites 3 and 2 were catalyzed by the same kinase. This multifunctional kinase was distinguished from glycogen synthase kinase-3 on the basis of nucleotide (ATP versus GTP) and protein substrate (glycogen synthase, ATP-citrate lyase, and acetyl-CoA carboxylase) specificities. Since the phosphate contents in glycogen synthase of sites 3 and 2 are altered in diabetes and by insulin administration, the possible involvement of the multifunctional kinase was explored. Glycogen synthase purified from diabetic rabbits was phosphorylated in vitro by this multifunctional kinase at only 10% of the rate compared to synthase purified from control rabbits. Treatment of the diabetics with insulin restored the synthase to a form that was readily phosphorylated in vitro.
mesh_terms: D000818:Animals; D001665:Binding Sites; D006006:Glycogen Synthase / Q000378:metabolism*; D007700:Kinetics; D008970:Molecular Weight; D009132:Muscles / Q000201:enzymology*; D010761:Phosphorus Radioisotopes; D010766:Phosphorylation; D011485:Protein Binding; D011494:Protein Kinases / Q000378:metabolism*; D011817:Rabbits; D013329:Structure-Activity Relationship
chemical_list: D010761:Phosphorus Radioisotopes; D006006:Glycogen Synthase; D011494:Protein Kinases; C032202:ATP-citrate lyase kinase