Mutations of muscle glycogen synthase that disable activation by glucose 6-phosphate

Isao Hanashiro, Peter Roach

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

Glycogen synthase, an enzyme of historical importance in the field of reversible protein modification, is inactivated by phosphorylation and allosterically activated by glucose 6-phosphate (glucose-6-P). Previous analysis of yeast glycogen synthase had identified a conserved and highly basic 13-amino-acid segment in which mutation of Arg residues resulted in loss of activation by glucose-6-P (12). The equivalent mutations R578R579R581A (all three of the indicated Arg residues mutated to Ala) and R585R587R590A were introduced into rabbit muscle glycogen synthase. Whether expressed transiently in COS-1 cells or produced in and purified from Escherichia coli, both mutant enzymes were insensitive to activation by glucose-6-P. The effect of phosphorylation was studied in two ways. Purified, recombinant glycogen synthase was directly phosphorylated by casein kinase 2 and glycogen synthase kinase 3, under conditions that inactivate the wild-type enzyme. In addition, phosphorylation sites were converted to Ala by mutagenesis in wild-type and in the glucose-6-P desensitized mutants expressed in COS-1 cells. Phosphorylation inactivated the R578R579R581A mutant but had little effect on the R585R587R590A. This result was surprising since phosphorylation had the opposite effects on the corresponding yeast enzyme mutants. The results confirm that the region of glycogen synthase, Arg.578-Arg-590, is required for activation by glucose-6-P and suggest that it is part of a sensitive and critical switch involved in transitions between different conformational states. However, the role must differ subtly between the mammalian and the yeast enzymes.

Original languageEnglish
Pages (from-to)286-292
Number of pages7
JournalArchives of Biochemistry and Biophysics
Volume397
Issue number2
DOIs
StatePublished - Jan 15 2002

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Glycogen Synthase
Glucose-6-Phosphate
Phosphorylation
Muscle
Chemical activation
Muscles
Mutation
Yeast
Enzymes
Yeasts
COS Cells
Glycogen Synthase Kinase 3
Casein Kinase II
Basic Amino Acids
Mutagenesis
Escherichia coli
Switches
Rabbits
Amino Acids
Glucose

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Mutations of muscle glycogen synthase that disable activation by glucose 6-phosphate. / Hanashiro, Isao; Roach, Peter.

In: Archives of Biochemistry and Biophysics, Vol. 397, No. 2, 15.01.2002, p. 286-292.

Research output: Contribution to journalArticle

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