Glycogen debranching enzyme in bovine brain.
Glycogen debranching enzyme was partially purified from bovine brain using a substrate for measuring the amylo-1,6-glucosidase activity. Bovine cerebrum was homogenized, followed by cell-fractionation of the resulting homogenate. The enzyme activity was found mainly in the cytosolic fraction. The enzyme was purified 5,000-fold by ammonium sulfate precipitation, anion-exchange chromatography, gel-filtration, anion-exchange HPLC, and gel-permeation HPLC. The enzyme preparation had no alpha-glucosidase or alpha-amylase activities and degraded phosphorylase limit dextrin of glycogen with phosphorylase. The molecular weight of the enzyme was 190,000 and the optimal pH was 6.0. The brain enzyme differed from glycogen debranching enzyme of liver or muscle in its mode of action on dextrins with an alpha-1,6-glucosyl branch, indicating an amino acid sequence different from those of the latter two enzymes. It is likely that the enzyme is involved in the breakdown of brain glycogen in concert with phosphorylase as in the cases of liver and muscle, but that this proceeds in a somewhat different manner. The enzyme activity decreased in the presence of ATP, suggesting that the degradation of brain glycogen is controlled by the modification of the debranching enzyme activity as well as the phosphorylase.
Narahara
E
E
Department of Environmental Sciences, Faculty of Science, Osaka Women's University, Sakai, Osaka 590-0035, Japan.
Makino
Y
Y
Omichi
K
K
D000249:Adenosine Monophosphate / Q000378:metabolism / Q000494:pharmacology; D000255:Adenosine Triphosphate / Q000378:metabolism / Q000494:pharmacology; D000494:Allosteric Regulation / Q000502:physiology; D000818:Animals; D001921:Brain / Q000201:enzymology*; D002417:Cattle; D005947:Glucose / Q000378:metabolism / Q000494:pharmacology; D006003:Glycogen / Q000378:metabolism*; D006004:Glycogen Debranching Enzyme System / Q000187:drug effects / Q000302:isolation & purification* / Q000378:metabolism*; D024981:Glycogen Phosphorylase / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D008099:Liver / Q000201:enzymology; D008670:Metals / Q000378:metabolism; D009132:Muscles / Q000201:enzymology; D013379:Substrate Specificity / Q000502:physiology
D006004:Glycogen Debranching Enzyme System; D008670:Metals; D000249:Adenosine Monophosphate; D000255:Adenosine Triphosphate; D006003:Glycogen; D024981:Glycogen Phosphorylase; C030563:amylo-1,6-glucosidase; D005947:Glucose