Glycogen phosphorylase in fish muscle: demonstration of three interconvertible forms.
White skeletal muscle of crucian carp contains a single isoenzyme of glycogen phosphorylase, which was purified approximately 300-fold to a specific activity of approximately 13 mumol.min-1.mg protein-1 (assayed in the direction of glycogen breakdown at 25 degrees C). Tissue extracts of crucian muscle produced three distinct peaks of phosphorylase activity when separated on DEAE-Sephacel. Peaks 1 and 3 were identified, in terms of kinetic properties and by interconversion experiments, as phosphorylase b and a, respectively. Peak 2 was shown to be a phospho-dephospho hybrid. The three interconvertible forms of phosphorylase were purified and shown to be dimeric molecules at 20 degrees C. At 5 degrees C, a and the hybrid tended to form tetramers. The Mr of the subunit was estimated to be 96,400 from sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The hybrid is kinetically homogeneous, and its kinetic properties are intermediate between those of b and a forms. The b, hybrid, and a forms of phosphorylase can be isolated from rapidly frozen muscle of crucian but in different proportions, depending on whether fish were anesthetized or forced to muscular activity for 20 s. Muscle of anesthetized crucian had 36, 36, and 28% of phosphorylase b, hybrid, and a forms, respectively, whereas the corresponding values for exercised fish were 12, 37, and 51%. Results suggest that three interconvertible forms of phosphorylase exist simultaneously in crucian muscle and that hybrid phosphorylase is active in contracting muscle in vivo.
Schmidt
H
H
Institut für Zoologie, Johannes Gutenberg-Universität Mainz, Federal Republic of Germany.
Wegener
G
G
D000768:Anesthesia, General; D000818:Animals; D002347:Carps; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D007527:Isoenzymes / Q000302:isolation & purification / Q000378:metabolism*; D007700:Kinetics; D009132:Muscles / Q000201:enzymology*; D010762:Phosphorylase a / Q000302:isolation & purification / Q000378:metabolism*; D010763:Phosphorylase b / Q000302:isolation & purification / Q000378:metabolism*; D006005:Phosphorylases / Q000378:metabolism*; D010766:Phosphorylation; D005082:Physical Exertion
D007527:Isoenzymes; D010762:Phosphorylase a; D010763:Phosphorylase b; D006005:Phosphorylases