Analytical gel chromatography of rabbit muscle amylo-1,6-glucosidase/4-alpha-glucanotransferase under denaturing and non-denaturing conditions.
Rabbit muscle amylo-1,6-glucosidase/4-alpha-glucanotransferase has a molecular weight of 160 000-170 000 as determined by analytical gel chromatography under denaturing and non-denaturing conditions. Three different data analyses of the results of gel chromatography in the presence of 6 M guanidine-HCl on 4% agarose give values of 159 000, 161 000, and 165 000 for amylo-1,6-glucosidase/4-alpha-glucanotransferase from rabbit muscle. Analytical gel chromatography under non-denaturing conditions gives a value of 170 000 for the molecular weight and a value of 4.8 nm for the Stokes'' radius. A calculation based on the Stokes radius using the Svedberg equation gives a value of 160 000 for the molecular weight. This verifies by two independent methods the previously reported molecular weight for the protein using sodium dodecyl sulphate-polyacrylamide gel electrophoresis. The results establish that the mammalian debranching enzyme is a single polypeptide chain containing two enzymatic activities.
D000818:Animals; D001665:Binding Sites; D002850:Chromatography, Gel; D005964:Glucosyltransferases / Q000302:isolation & purification; D006026:Glycoside Hydrolases / Q000302:isolation & purification; D006146:Guanidines; D046911:Macromolecular Substances; D008970:Molecular Weight; D009132:Muscles / Q000201:enzymology*; D011485:Protein Binding; D011489:Protein Denaturation; D011817:Rabbits
D006146:Guanidines; D046911:Macromolecular Substances; D005964:Glucosyltransferases; D006026:Glycoside Hydrolases