title: Purification of a new high activity form of glucose-6-phosphate dehydrogenase from rat liver and the effect of enzyme inactivation on its immunochemical reactivity.
abstract: A new form of cytoplasmic glucose-6-phosphate dehydrogenase (E.C.1.1.1.49) was purified from rat liver by protamine sulfate precipitation, ammonium sulfate fractionation, ion exchange chromatography with diethylaminoethyl cellulose, and affinity chromatography with Cibacron blue agarose and NADP agarose. This form of the enzyme has a specific activity of over 600 units/mg of protein and gives essentially a single band by polyacrylamide gel electrophoresis. The form of the enzyme isolated by this purification method is 3 times more active than the form purified from liver by previously reported procedures. The relative mass of this pure glucose-6-phosphate dehydrogenase enzyme was determined by disc gel electrophoresis to be 269,000. This high activity glucose-6-phosphate dehydrogenase enzyme, after inactivation by reaction with palmityl-CoA, was no longer precipitated by specific rabbit and goat antisera to this purified enzyme. Thus, the possibility still exists that starved fat-refed animals contain glucose-6-phosphate dehydrogenase (G6PD) enzyme protein in an inactivated form no longer detectable by either enzyme activity or immunoprecipitation.
mesh_terms: D000818:Animals; D000939:Epitopes; D005954:Glucosephosphate Dehydrogenase / Q000037:antagonists & inhibitors / Q000276:immunology / Q000302:isolation & purification*; D006041:Goats / Q000276:immunology; D007106:Immune Sera; D005779:Immunodiffusion; D007122:Immunoelectrophoresis; D007123:Immunoelectrophoresis, Two-Dimensional; D008099:Liver / Q000201:enzymology*; D008297:Male; D008970:Molecular Weight; D051381:Rats
chemical_list: D000939:Epitopes; D007106:Immune Sera; D005954:Glucosephosphate Dehydrogenase