Phosphorylation of myosin light chain kinase by the multifunctional calmodulin-dependent protein kinase II in smooth muscle cells

Malú G. Tansey, R. Ann Word, Hiroyoshi Hidaka, Harold A. Singer, Charles M. Schworer, Kristine E. Kamm, James T. Stull

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Abstract

Stimulation of tracheal smooth muscle cells in culture with ionomycin resulted in a rapid increase in cytosolic free Ca2+ concentration ([Ca2+]i) and an increase in both myosin light chain kinase and myosin light chain phosphorylation. These responses were markedly inhibited in the absence of extracellular Ca2+. Pretreatment of cells with 1-[N-O-bis(5-isoquinolinesulfonyl)-N-methyl-L-tyrosyl]-4-phenylpiperazine (KN-62), a specific inhibitor of the multifunctional calmodulin-dependent protein kinase II (CaM kinase II), did not affect the increase in [Ca2+]i but inhibited ionomycin-induced phosphorylation of myosin light chain kinase at the regulatory site near the calmodulin-binding domain. KN-62 inhibited CaM kinase II activity toward purified myosin light chain kinase. Phosphorylation of myosin light chain kinase decreased its sensitivity to activation by Ca2+ in cell lysates. Pretreatment of cells with KN-62 prevented this desensitization to Ca2+ and potentiated myosin light chain phosphorylation. We propose that the Ca2+-dependent phosphorylation of myosin light chain kinase by CaM kinase II decreases the Ca2+ sensitivity of myosin light chain phosphorylation in smooth muscle.

Original languageEnglish (US)
Pages (from-to)12511-12516
Number of pages6
JournalJournal of Biological Chemistry
Volume267
Issue number18
StatePublished - Jun 25 1992

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KN 62
Myosin-Light-Chain Kinase
Calcium-Calmodulin-Dependent Protein Kinase Type 2
Phosphorylation
Smooth Muscle Myocytes
Muscle
Cells
Myosin Light Chains
Ionomycin
Calmodulin
Cell culture
Smooth Muscle
Cell Culture Techniques
Chemical activation

ASJC Scopus subject areas

  • Biochemistry

Cite this

Phosphorylation of myosin light chain kinase by the multifunctional calmodulin-dependent protein kinase II in smooth muscle cells. / Tansey, Malú G.; Word, R. Ann; Hidaka, Hiroyoshi; Singer, Harold A.; Schworer, Charles M.; Kamm, Kristine E.; Stull, James T.

In: Journal of Biological Chemistry, Vol. 267, No. 18, 25.06.1992, p. 12511-12516.

Research output: Contribution to journalArticle

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AU - Schworer, Charles M.

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AU - Stull, James T.

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