Activation of cGMP-dependent protein kinase by protein kinase C

Yali Hou, Judith Lascola, Nickolai O. Dulin, Richard D. Ye, Darren D Browning

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

The cGMP-dependent protein kinases (PKG) are emerging as important components of mainstream signal transduction pathways. Nitric oxide-induced cGMP formation by stimulation of soluble guanylate cyclase is generally accepted as being the most widespread mechanism underlying PKG activation. In the present study, PKG was found to be a target for phorbol 12-myristate 13-acetate (PMA)-responsive protein kinase C (PKC). PKG1α became phosphorylated in HEK-293 cells stimulated with PMA and also in vitro using purified components. PKC-dependent phosphorylation was found to activate PKG as measured by phosphorylation of vasodilator-stimulated phosphoprotein, and by in vitro kinase assays. Although there are 11 potential PKC substrate recognition sites in PKG1α, threonine 58 was examined due to its proximity to the pseudosubstrate domain. Antibodies generated against the phosphorylated form of this region were used to demonstrate phosphorylation in response to PMA treatment of the cells with kinetics similar to vasodilator-stimulated phosphoprotein phosphorylation. A phospho-mimetic mutation at this site (T58E) generated a partially activated PKG that was more sensitive to cGMP levels. A phospho-null mutation (T58A) revealed that this residue is important but not sufficient for PKG activation by PKC. Taken together, these findings outline a novel signal transduction pathway that links PKC stimulation with cyclic nucleotide-independent activation of PKG.

Original languageEnglish (US)
Pages (from-to)16706-16712
Number of pages7
JournalJournal of Biological Chemistry
Volume278
Issue number19
DOIs
StatePublished - May 9 2003

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Cyclic GMP-Dependent Protein Kinases
Phosphorylation
Protein Kinase C
Chemical activation
Signal transduction
Acetates
Signal Transduction
Mutation
HEK293 Cells
Guanylate Cyclase
Cyclic Nucleotides
Threonine
Assays
Nitric Oxide
Phosphotransferases
Cells
Kinetics
Antibodies
Substrates
phorbol-12-myristate

ASJC Scopus subject areas

  • Biochemistry

Cite this

Activation of cGMP-dependent protein kinase by protein kinase C. / Hou, Yali; Lascola, Judith; Dulin, Nickolai O.; Ye, Richard D.; Browning, Darren D.

In: Journal of Biological Chemistry, Vol. 278, No. 19, 09.05.2003, p. 16706-16712.

Research output: Contribution to journalArticle

Hou, Yali ; Lascola, Judith ; Dulin, Nickolai O. ; Ye, Richard D. ; Browning, Darren D. / Activation of cGMP-dependent protein kinase by protein kinase C. In: Journal of Biological Chemistry. 2003 ; Vol. 278, No. 19. pp. 16706-16712.
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