Phosphorylation site specificities of glycogen synthase kinases: Determination by peptide mapping using high-performance liquid chromatography

Henning Juhl, Virender S. Sheorain, Charles M. Schworer, Mary Frances Jett, Thomas Soderling

Research output: Contribution to journalArticle

20 Citations (Scopus)

Abstract

A method is described which separates the various phosphorylation sites in glycogen synthase based on reverse phase high-performance liquid chromatography (HPLC) of tryptic 32P-peptides. Using this method we studied the phosphorylation site specificities of the kinases which act on glycogen synthase. The cAMP-dependent protein kinase phosphorylated sites 1a, 1b, and 2, whereas casein kinase II phosphorylated only site 5. Two calcium, calmodulin-dependent kinases, phosphorylase kinase and liver calmodulin-dependent synthase kinase, both phosphorylated site 2, and the latter enzyme also phosphorylated site 1b. A cAMP-independent kinase (kinase 4) purified from liver also specifically phosphorylated site 2. Synthase kinase 3 catalyzed the phosphorylation of only site 3. This HPLC method was also used to establish that all of these sites were subject to phosphorylation in vivo.

Original languageEnglish (US)
Pages (from-to)518-526
Number of pages9
JournalArchives of Biochemistry and Biophysics
Volume222
Issue number2
DOIs
StatePublished - Apr 15 1983
Externally publishedYes

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Glycogen Synthase Kinases
Phosphorylation
Peptide Mapping
High performance liquid chromatography
Phosphotransferases
High Pressure Liquid Chromatography
Peptides
Glycogen Synthase
Calmodulin
Liver
Calcium-Calmodulin-Dependent Protein Kinase Kinase
Phosphorylase Kinase
Casein Kinase II
Reverse-Phase Chromatography
Cyclic AMP-Dependent Protein Kinases
Calcium
Enzymes

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Phosphorylation site specificities of glycogen synthase kinases : Determination by peptide mapping using high-performance liquid chromatography. / Juhl, Henning; Sheorain, Virender S.; Schworer, Charles M.; Jett, Mary Frances; Soderling, Thomas.

In: Archives of Biochemistry and Biophysics, Vol. 222, No. 2, 15.04.1983, p. 518-526.

Research output: Contribution to journalArticle

Juhl, Henning ; Sheorain, Virender S. ; Schworer, Charles M. ; Jett, Mary Frances ; Soderling, Thomas. / Phosphorylation site specificities of glycogen synthase kinases : Determination by peptide mapping using high-performance liquid chromatography. In: Archives of Biochemistry and Biophysics. 1983 ; Vol. 222, No. 2. pp. 518-526.
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