Endogenous phosphates on liver glycogen synthase D and synthase I. Studies on the number and location.
Synthase D and synthase I of high specific activity have been isolated from rabbit liver, with subunit Mr = 78,000 and 85,000, respectively. Minor bands of Mr = 78,000 and 71,000 were also observed, and these were shown to be proteolytic products containing synthase activity. Incubation with trypsin resulted in no activity changes, but there was a conversion of synthase I to the Mr = 78,000 species. Incubation with chymotrypsin produced no change in total synthase activity, but both enzymes were converted to the Mr = 71,000 species. Synthase I, after chymotrypsin treatment, became dependent on glucose-6-P for activity. Kinetic studies indicated that the chymotrypsin-treated synthase D and synthase I were not the same species. Synthase D contained 5.7 mol of alkali-labile phosphate/subunit and synthase I, 2.9 mol/subunit. With trypsin treatment there was little loss of phosphate from synthase. With chymotrypsin treatment, there was a loss of 3.1 and 1.5 mol of phosphate/subunit from synthase D and I, respectively. Some conversion to synthase I still occurred when chymotrypsin-treated synthase D was incubated with crude extract, suggesting that the phosphates which remained were still removable by synthase phosphatase. The presence of additional alkali-stable phosphates was indicated when the synthase samples were ashed. The total phosphates were 17.3 mol/subunit for synthase D and 13.4 mol/subunit for synthase I. Our results indicate that the liver synthase enzymes are highly phosphorylated and extremely susceptible to endogenous and exogenous proteolysis. The alkali-labile phosphates are distributed in chymotrypsin-sensitive as well as chymotrypsin-insensitive sites. Both sites are involved in the conversion of synthase D to synthase I.
D000818:Animals; D002918:Chymotrypsin / Q000378:metabolism; D006006:Glycogen Synthase / Q000032:analysis*; D006007:Glycogen-Synthase-D Phosphatase / Q000378:metabolism; D007527:Isoenzymes / Q000032:analysis; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D008297:Male; D008970:Molecular Weight; D010710:Phosphates / Q000032:analysis*; D011817:Rabbits; D014357:Trypsin / Q000378:metabolism
D007527:Isoenzymes; D010710:Phosphates; D006006:Glycogen Synthase; D006007:Glycogen-Synthase-D Phosphatase; D002918:Chymotrypsin; D014357:Trypsin