Phosphorylation of smooth muscle myosin heavy and light chains. Effects of phorbol dibutyrate and agonists

K. E. Kamm, L. C. Hsu, Y. Kubota, J. T. Stull

Research output: Contribution to journalArticle

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Abstract

A number of different protein kinases phosphorylate purified heavy chains or the 20-kDa light chain of smooth muscle myosin. The physiological significance of these phosphorylation reactions has been examined in intact smooth muscle. Myosin heavy chain was slightly phosphorylated (0.08 mol of phosphate/mol) under control conditions in bovine tracheal tissue. Treatment with carbachol, isoproterenol, or phorbol 12,13-dibutyrate resulted in no significant change. In contrast, heavy chain was phorphorylated to 0.30 mol of phosphate/mol of heavy chain in tracheal smooth muscle cells in culture. This value increased significantly with ionomycin treatment. In control tissues, 9% of the light chain was monophosphorylated with 32P in the serine site phosphorylated by myosin light chain kinase. Carbachol (0.1 μM) alone resulted in contraction and 42% monophosphorylated light chain with 32P only in the serine site phosphorylated by myosin light chain kinase. Similarly, stimulation with histamine, 5-hydroxytryptamine, or KCl resulted in 32P incorporation into only the myosin light chain kinase serine site. Phorbol 12,13-dibutyrate (1 μM) alone resulted in 22% monophosphorylated light chain. However, only 25% of the 32P was in the myosin light chain kinase serine site, whereas 75% was in a serine site phosphorylated by protein kinase C. Phorbol 12,13-dibutyrate plus carbachol resulted in 27% monophosphorylated light chain; 75% of the 32P was in the myosin light chain kinase serine site, with the remainder in the protein kinase C serine site. These results indicate that phorbol esters act to increase phosphorylation of myosin light chain by protein kinase C. However, receptor-mediated stimulation or depolarization leading to tracheal smooth muscle contraction results in phosphorylation of myosin light chain by myosin light chain kinase alone.

Original languageEnglish (US)
Pages (from-to)21223-21229
Number of pages7
JournalJournal of Biological Chemistry
Volume264
Issue number35
StatePublished - 1989

Fingerprint

Smooth Muscle Myosins
Myosin-Light-Chain Kinase
Myosin Light Chains
Phosphorylation
Myosin Heavy Chains
Serine
Phorbol 12,13-Dibutyrate
Light
Carbachol
Muscle
Protein Kinase C
Smooth Muscle
Phosphates
Tissue
Ionomycin
Depolarization
Phorbol Esters
Muscle Contraction
phorbol
Isoproterenol

ASJC Scopus subject areas

  • Biochemistry

Cite this

Phosphorylation of smooth muscle myosin heavy and light chains. Effects of phorbol dibutyrate and agonists. / Kamm, K. E.; Hsu, L. C.; Kubota, Y.; Stull, J. T.

In: Journal of Biological Chemistry, Vol. 264, No. 35, 1989, p. 21223-21229.

Research output: Contribution to journalArticle

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