Glucose-stimulated phosphorylation of the 64-kDa protein of human polymorphonuclear leukocytes in a cell-free system

Glucose-stimulated phosphorylation of 64-kDa protein using a greater than 30 kDa fraction of human polymorphonuclear leukocytes in a dose-dependent fashion with 33 microM for maximum stimulation and 1.4 microM for ED50. None of the glucose derivatives and metabolites of glycolysis stimulated phosphorylation, but glucose-1-phosphate, glucose-6-phosphate, fructose-6-phosphate and fructose-1,6-diphosphate inhibited the glucose-stimulated phosphorylation, strongly suggesting phosphorylation by glucose-regulated protein kinase. Cyclic nucleotides and the protein kinase inhibitors H-7, H-8, W-7 and staurosporine did not affect phosphorylation.