Studies on the effect of phosphorylation of the 20,000 Mr light chain of vertebrate smooth muscle myosin

J. Kendrick-Jones, W. Z. Cande, P. J. Tooth, R. C. Smith, J. M. Scholey

Research output: Contribution to journalArticle

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Abstract

Myosin was rapidly prepared from turkey gizzard muscle to a high level of purity, in high yield and in a non-phosphorylated state. It was consistently observed that the actin-activated Mg2+ ATPase activity of this myosin was dependent on the level of phosphorylation of the 20,000 Mr light chain, for example, in the non-phosphorylated state, the myosin Mg2+ ATPase activity was not activated by actin whereas, when the light chains were phosphorylated, the Mg2+ ATPase activity of the myosin was activated approximately ninefold by actin. Using the "desensitized" scallop myosin test system (Kendrick-Jones et al., 1976; Sellers et al., 1980) it was further demonstrated that phosphorylation of the 20,000 Mr gizzard light chain has a regulatory role. These results also suggest that the regulatory mechanisms mediated by smooth muscle myosin light chains and molluscan myosin regulatory light chains are similar, i.e. in the absence of Ca2+, both types of light chain inhibit myosin interaction with actin and this inhibition is relieved by either phosphorylation in smooth muscle or by direct calcium binding in molluscan myosins. The basis of regulation exerted by these light chains is therefore repression-derepression. Using a variety of techniques, i.e. turbidity measurements, quantitative high speed centrifugation, electron microscopy and dark field light microscopy, it was observed that the stability of gizzard myosin filaments at approximately physiological conditions (0·15 m-NaCl, 1 mm-MgATP, pH 7·0) was dependent on the level of light chain phosphorylation. Using purified calmodulin-dependent light chain kinase and phosphatase, it was further shown that these gizzard myosin filaments can be reversibly assembled and disassembled as a result of phosphorylation-dephosphorylation of the 20,000 Mr light chain.

Original languageEnglish (US)
Pages (from-to)139-162
Number of pages24
JournalJournal of Molecular Biology
Volume165
Issue number1
DOIs
StatePublished - Mar 25 1983
Externally publishedYes

Fingerprint

Smooth Muscle Myosins
Myosins
Vertebrates
Phosphorylation
Light
Ca(2+) Mg(2+)-ATPase
Myosin Light Chains
Actins
Pectinidae
Calmodulin
Centrifugation
Phosphoric Monoester Hydrolases
Smooth Muscle
Microscopy
Electron Microscopy
Phosphotransferases
Adenosine Triphosphate
Calcium
Muscles

ASJC Scopus subject areas

  • Molecular Biology
  • Virology

Cite this

Studies on the effect of phosphorylation of the 20,000 Mr light chain of vertebrate smooth muscle myosin. / Kendrick-Jones, J.; Cande, W. Z.; Tooth, P. J.; Smith, R. C.; Scholey, J. M.

In: Journal of Molecular Biology, Vol. 165, No. 1, 25.03.1983, p. 139-162.

Research output: Contribution to journalArticle

Kendrick-Jones, J. ; Cande, W. Z. ; Tooth, P. J. ; Smith, R. C. ; Scholey, J. M. / Studies on the effect of phosphorylation of the 20,000 Mr light chain of vertebrate smooth muscle myosin. In: Journal of Molecular Biology. 1983 ; Vol. 165, No. 1. pp. 139-162.
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