Activation of 4-alpha-glucanotransferase activity of porcine liver glycogen debranching enzyme with cyclodextrins.
Glycogen debranching enzyme (GDE) is a single polypeptide chain containing distinct active sites for 4-alpha-glucanotransferase and amylo-alpha-1,6-glucosidase activities. Debranching of phosphorylase limit dextrin from glycogen is carried out by cooperation of the two activities. We examined the effects of cyclodextrins (CDs) on debranching activity of porcine liver GDE using a fluorogenic branched dextrin, Glcalpha1-4Glcalpha1-4Glcalpha1-4(Glcalpha1-4Glcalpha1-4Glcalpha1-4Glcalpha1-6)Glcalpha1-4Glcalpha1-4Glcalpha1-4Glcalpha1-4GlcPA (B5/84), as a substrate. B5/84 was hydrolyzed by the hydrolytic action of 4-alpha-glucanotransferase to B5/81 and maltotriose. The fluorogenic product was further hydrolyzed by the amylo-alpha-1,6-glucosidase activity to the debranched product, Glcalpha1-4Glcalpha1-4Glcalpha1-4Glcalpha1-4Glcalpha1-4Glcalpha1-4Glcalpha1-4GlcPA (G8PA), and glucose. alpha-, beta- and gamma-CDs accelerated the liberation of B5/81 from B5/84, indicating that the 4-alpha-glucanotransferase activity was activated by CDs to remove the maltotriosyl residue from the maltotetraosyl branch. This led to acceleration of B5/84 debranching. The extent of 4-alpha-glucanotransferase activation increased with CD concentration before reaching a constant value. This suggests that there is an activator binding site and that the binding of CDs stimulates 4-alpha-glucanotransferase activity. In the porcine liver, glycogen degradation may be partially stimulated by the binding of a glycogen branch to this activator binding site.
Watanabe
Yumiko
Y
Department of Chemistry, Graduate School of Science, Osaka Prefecture University, 2-1 Daisen-cho, Sakai, Osaka 590-0035.
Makino
Yasushi
Y
Omichi
Kaoru
K
D000818:Animals; D001665:Binding Sites; D002240:Carbohydrate Sequence; D002851:Chromatography, High Pressure Liquid; D003505:Cyclodextrins / Q000494:pharmacology*; D004789:Enzyme Activation; D006004:Glycogen Debranching Enzyme System / Q000302:isolation & purification / Q000378:metabolism*; D008099:Liver / Q000201:enzymology*; D008969:Molecular Sequence Data; D013552:Swine
D003505:Cyclodextrins; D006004:Glycogen Debranching Enzyme System; C020569:4 alpha-glucanotransferase