Glycogenin is the priming glucosyltransferase required for the initiation of glycogen biogenesis in rabbit skeletal muscle.
Purified preparations of glycogen synthase are a complex of two proteins, the catalytic subunit of glycogen synthase and glycogenin, present in a 1:1 molar ratio [J. Pitcher, C. Smythe, D. G. Campbell & P. Cohen (1987) Eur. J. Biochem. 169, 497-502]. This complex has now been found to contain a further glucosyltransferase activity that catalyses the transfer of glucose residues from UDP-Glc to glucosylated-glycogenin. The glucosyltransferase, which is of critical importance in forming the primer required for de novo glycogen biosynthesis, is distinct from glycogen synthase in several ways. It has an absolute requirement for divalent cations, a 1000-fold lower Km for UDP-Glc and its activity is unaffected by incubation with UDP-pyridoxal or exposure to 2 M LiBr, which inactivate glycogen synthase by 95% and 100%, respectively. The priming glucosyltransferase and glycogen synthase activities coelute on Superose 6, and the rate of glycosylation of glycogenin is independent of enzyme concentration, suggesting that the reaction is catalysed intramolecularly by a subunit of the glycogen synthase complex. This component has been identified as glycogenin, following dissociation of the subunits in 2 M LiBr and their separation on Superose 12. The glycosylation of isolated glycogenin reaches a plateau when five additional glucose residues have been added to the protein, and digestion with alpha-amylase indicates that all the glycogenin molecules contain at least one glucosyl residue prior to autoglucosylation. The priming glucosyltransferase activity of glycogenin is unaffected by either glucose 6-phosphate or by phosphorylation of the catalytic subunit of glycogen synthase. The mechanism of primer formation is discussed in the light of the finding that glycogenin is an enzyme that catalyses its own autoglucosylation.
Pitcher
J
J
Department of Biochemistry, University of Dundee, Scotland.
Smythe
C
C
Cohen
P
P
D000818:Animals; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D005964:Glucosyltransferases / Q000378:metabolism*; D006003:Glycogen / Q000096:biosynthesis*; D006006:Glycogen Synthase / Q000378:metabolism; D006023:Glycoproteins / Q000378:metabolism*; D009132:Muscles / Q000201:enzymology / Q000378:metabolism*; D011817:Rabbits
D006023:Glycoproteins; C051887:glycogenin; D006003:Glycogen; D005964:Glucosyltransferases; D006006:Glycogen Synthase