Rat heart glycogen phosphorylase II is genetically distinct from phosphorylase I.
Previous studies in our laboratory have shown that rat heart glycogen phosphorylase (1,4-alpha-D-glucan: orthophosphate alpha-D-glucosyltransferase, EC 2.4.1.1) separates into two forms upon ion-exchange chromatography. Both forms could be shown to have the same subunit Mr and to incorporate one molecule of phosphate per subunit. The studies reported here were done to check whether both forms are native isoenzymes and, further, which form might represent the heart-specific phosphorylase. Firstly, the iso-electric points of the purified enzymes are compared with those associated with phosphorylase activity in crude extracts from rat heart. Two out of four major bands coincided with the bands of purified phosphorylase Ib and IIb (isoelectric points: 5.5 and 6.25), indicating apparent identity. Secondly, antibodies to rat skeletal muscle phosphorylase reacted with rat heart phosphorylase I, whereas phosphorylase II was neither inhibited nor precipitated by the antibody. Thirdly, peptide maps obtained after proteolytic digestion of SDS-denatured phosphorylase I and II showed different patterns. In addition to the kinetic differences between these two forms reported earlier, phosphorylase IIa was inhibited by glucose 6-phosphate, whereas phosphorylase Ia was not. These results suggest that phosphorylase II is a heart-specific isoenzyme which is presumably encoded by a different gene.
D000818:Animals; D005779:Immunodiffusion; D007122:Immunoelectrophoresis; D007158:Immunologic Techniques; D007525:Isoelectric Focusing; D007527:Isoenzymes / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D008297:Male; D009132:Muscles / Q000201:enzymology; D009206:Myocardium / Q000201:enzymology*; D010446:Peptide Fragments; D010447:Peptide Hydrolases / Q000378:metabolism; D006005:Phosphorylases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D010766:Phosphorylation; D051381:Rats; D011919:Rats, Inbred Strains; D014357:Trypsin / Q000378:metabolism
D007527:Isoenzymes; D010446:Peptide Fragments; D006005:Phosphorylases; D010447:Peptide Hydrolases; D014357:Trypsin