Phosphorylation of sites 3 and 2 in rabbit skeletal muscle glycogen synthase by a multifunctional protein kinase (ATP-citrate lyase kinase)

V. S. Sheorain, S. Ramakrishna, W. B. Benjamin, Thomas Soderling

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

A multifunctional protein kinase, purified from rat liver as ATP-citrate lyase kinase, has been identified as a glycogen synthase kinase. This kinase catalyzed incorporation of up to 1.5 mol of 32PO4/mol of synthase subunit associated with a decrease in the glycogen synthase activity ratio from 0.85 to a value of 0.15. Approximately 65-70% of the 32PO4 was incorporated into site 3 and 30-35% into site 2 as determined by reverse phase high performance liquid chromatography. Release of 32PO4 from the phosphopeptides during automated Edman degradation confirmed the site 3 and 2 assignment. Thermal stability studies established that the phosphorylations of sites 3 and 2 were catalyzed by the same kinase. This multifunctional kinase was distinguished from glycogen synthase kinase-3 on the basis of nucleotide (ATP versus GTP) and protein substrate (glycogen synthase, ATP-citrate lyase, and acetyl-CoA carboxylase) specificities. Since the phosphate contents in glycogen synthase of sites 3 and 2 are altered in diabetes and by insulin administration, the possible involvement of the multifunctional kinase was explored. Glycogen synthase purified from diabetic rabbits was phosphorylated in vitro by this multifunctional kinase at only 10% of the rate compared to synthase purified from control rabbits. Treatment of the diabetics with insulin restored the synthase to a form that was readily phosphorylated in vitro.

Original languageEnglish (US)
Pages (from-to)12287-12292
Number of pages6
JournalJournal of Biological Chemistry
Volume260
Issue number22
StatePublished - 1985
Externally publishedYes

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Glycogen Synthase
Phosphorylation
Protein Kinases
Muscle
Skeletal Muscle
Phosphotransferases
Rabbits
ATP Citrate (pro-S)-Lyase
Insulin
Glycogen Synthase Kinase 3
Acetyl-CoA Carboxylase
Glycogen Synthase Kinases
Phosphopeptides
High performance liquid chromatography
Reverse-Phase Chromatography
Medical problems
Guanosine Triphosphate
Liver
Rats
Thermodynamic stability

ASJC Scopus subject areas

  • Biochemistry

Cite this

Phosphorylation of sites 3 and 2 in rabbit skeletal muscle glycogen synthase by a multifunctional protein kinase (ATP-citrate lyase kinase). / Sheorain, V. S.; Ramakrishna, S.; Benjamin, W. B.; Soderling, Thomas.

In: Journal of Biological Chemistry, Vol. 260, No. 22, 1985, p. 12287-12292.

Research output: Contribution to journalArticle

Sheorain, V. S. ; Ramakrishna, S. ; Benjamin, W. B. ; Soderling, Thomas. / Phosphorylation of sites 3 and 2 in rabbit skeletal muscle glycogen synthase by a multifunctional protein kinase (ATP-citrate lyase kinase). In: Journal of Biological Chemistry. 1985 ; Vol. 260, No. 22. pp. 12287-12292.
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