Effect of metabolites and phosphorylase on the D to I conversion of glycogen synthase from human polymorphonuclear leukocytes.
The D to I conversion of glycogen synthase from human polymorphonuclear leukocytes was examined both in a gel-filtered homogenate and in a preparation of glycogen particles with adhering enzymes, purified by chromatography on concanavalin A bound to Sepharose. It was found that glucose 6-phosphate as well as mannose 6-phosphate, glucosamine 6-phosphate, and 2-deoxy-glucose 6-phosphate activated the reaction, whereas the corresponding sugars were without effect. Mn2+ and Ca2+ increased the conversion rate by 51% and 27%, respectively, whereas Mg2+ and inorganic phosphate were without effect. Sodium fluoride inhibited the reaction completely. Glycogen inhibited the reaction in physiological concentrations and 0.5 mM glucose 6-phosphate was able to overcome this inhibition. MgATP greatly augmented the inhibition caused by glycogen in the glycogen particle preparation. This combined effect could be overcome by glucose 6-phosphate in concentrations from 0.1 to 1 mM. Phosphorylase alpha purified from human polymorphonuclear leukocytes inhibited the D to I conversion in a glycogen particle preparation. The inhibition was counteracted by glucose 6-phosphate and to a lesser degree by AMP. Phosphorylase beta was also inhibitory, but only at higher concentrations than phosphorylase alpha. No phosphorylase phosphatase activity was found in the glycogen particle preparation, which may indicate that chromatography on concanavalin A-Sepharose separates this enzyme from the synthase phosphatase or partially destroys the activity of a hypothetical common protein phosphatase.
D000249:Adenosine Monophosphate / Q000494:pharmacology; D000255:Adenosine Triphosphate / Q000494:pharmacology; D002413:Cations, Divalent / Q000494:pharmacology; D000242:Cyclic AMP / Q000494:pharmacology; D005459:Fluorides / Q000494:pharmacology; D006003:Glycogen / Q000494:pharmacology; D006006:Glycogen Synthase / Q000097:blood*; D006600:Hexosephosphates / Q000494:pharmacology; D006601:Hexoses / Q000494:pharmacology; D006801:Humans; D008274:Magnesium / Q000494:pharmacology; D009504:Neutrophils / Q000201:enzymology*; D010710:Phosphates / Q000494:pharmacology; D006005:Phosphorylases / Q000494:pharmacology*
D002413:Cations, Divalent; D006600:Hexosephosphates; D006601:Hexoses; D010710:Phosphates; D000249:Adenosine Monophosphate; D000255:Adenosine Triphosphate; D006003:Glycogen; D000242:Cyclic AMP; D006005:Phosphorylases; D006006:Glycogen Synthase; D008274:Magnesium; D005459:Fluorides