Phosphate groups as substrate determinants for casein kinase I action

Horst Flotow, Paul R. Graves, Aiqun Wang, Carol J. Fiol, Roger W. Roeske, Peter Roach

Research output: Contribution to journalArticle

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Abstract

Phosphorylation of rabbit muscle glycogen synthase by cyclic AMP-dependent protein kinase has been shown to enhance subsequent phosphorylation by casein kinase I (Flotow, H., and Roach, P. J. (1989) J. Biol. Chem. 264, 9126-9128). In the present study, synthetic peptides based on the sequences of the four phosphorylated regions in muscle glycogen synthase were used to probe the role of substrate phosphorylation in casein kinase I action. With all four peptides, prior phosphorylation significantly stimulated phosphorylation by casein kinase I. A series of peptides was synthesized based on the NH2-terminal glycogen synthase sequence PLSRTLS7VSS10LPGL, in which phosphorylation at Ser7 is required for modification of Ser10 by casein kinase I. The spacing between the P-Ser and the acceptor Ser was varied to have 1, 2, or 3 intervening residues. The peptide with a 2-residue spacing (-S(P)-X-X-S-) was by far the best casein kinase I substrate. When the P-Ser residue at Ser was replaced with P-Thr, the resulting peptide was still a casein kinase I substrate. However, substitution of Asp or Glu residues at Ser7 led to peptides that were not phosphorylated by casein kinase I. Phosphorylation of one of the other peptides showed that Thr could also be the phosphate acceptor. From these results, we propose that there are substrates for casein kinase I for which prior phosphorylation is a critical determinant of protein kinase action. In these instances, an important recognition motif for casein kinase I appears to be -S(P)/T(P)-Xn-S/T- with n = 2 much more effective than n = 1 or n = 3. Thus, casein kinase I may be involved in hierarchal substrate phosphorylation schemes in which its activity is controlled by the phosphorylation state of its substrates.

Original languageEnglish
Pages (from-to)14264-14269
Number of pages6
JournalJournal of Biological Chemistry
Volume265
Issue number24
StatePublished - Aug 25 1990

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Casein Kinase I
Phosphorylation
Phosphates
Substrates
Peptides
Glycogen Synthase
Muscle
Muscles
Cyclic AMP-Dependent Protein Kinases
Protein Kinases
Substitution reactions

ASJC Scopus subject areas

  • Biochemistry

Cite this

Flotow, H., Graves, P. R., Wang, A., Fiol, C. J., Roeske, R. W., & Roach, P. (1990). Phosphate groups as substrate determinants for casein kinase I action. Journal of Biological Chemistry, 265(24), 14264-14269.

Phosphate groups as substrate determinants for casein kinase I action. / Flotow, Horst; Graves, Paul R.; Wang, Aiqun; Fiol, Carol J.; Roeske, Roger W.; Roach, Peter.

In: Journal of Biological Chemistry, Vol. 265, No. 24, 25.08.1990, p. 14264-14269.

Research output: Contribution to journalArticle

Flotow, H, Graves, PR, Wang, A, Fiol, CJ, Roeske, RW & Roach, P 1990, 'Phosphate groups as substrate determinants for casein kinase I action', Journal of Biological Chemistry, vol. 265, no. 24, pp. 14264-14269.
Flotow H, Graves PR, Wang A, Fiol CJ, Roeske RW, Roach P. Phosphate groups as substrate determinants for casein kinase I action. Journal of Biological Chemistry. 1990 Aug 25;265(24):14264-14269.
Flotow, Horst ; Graves, Paul R. ; Wang, Aiqun ; Fiol, Carol J. ; Roeske, Roger W. ; Roach, Peter. / Phosphate groups as substrate determinants for casein kinase I action. In: Journal of Biological Chemistry. 1990 ; Vol. 265, No. 24. pp. 14264-14269.
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