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PMID: 2531749 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Filamentous smooth muscle myosin is regulated by phosphorylation.

The Journal of cell biology ·Vol. 109 ·No. 6 Pt 1 ·1989-12-00 ·Pages 2887-94

Trybus KM

Abstract

The enzymatic activity of filamentous dephosphorylated smooth muscle myosin has been difficult to determine because the polymer disassembles to the folded conformation in the presence of MgATP. Monoclonal antirod antibodies were used here to "fix" dephosphorylated myosin in the filamentous state. The steady-state actin-activated ATPase of phosphorylated filaments was 30-100-fold higher than that of antibody-stabilized dephosphorylated filaments, suggesting that phosphorylation can activate ATPase activity independent of changes in assembly. The degree of regulation may exceed 100-fold, because steady-state measurements slightly overestimate the rate of product release from dephosphorylated filaments. Single-turnover experiments in the absence of actin showed that although dephosphorylated folded myosin released products at the low rate of 0.0005 s-1 (Cross, R. A., K. E. Cross, A. Sobieszek. 1986. EMBO [Eur. Mol. Biol. Organ.] J. 5:2637-2641) the rate of product release from dephosphorylated filaments was only 3-12-fold higher, depending on the ionic strength. The addition of actin did not increase this rate to any appreciable extent. Dephosphorylated filaments and dephosphorylated heavy meromyosin (Sellers, J. R. 1985. J. Biol. Chem. 260:15815-15819) thus have similar low rates of phosphate release both in the presence and absence of actin. These results show that light chain phosphorylation alone, without invoking other mechanisms, is an effective switch for regulating the activity of smooth muscle myosin filaments.

MeSH Terms
Actin Cytoskeleton/metabolism,ultrastructure Actins/metabolism Animals Enzyme Activation Gizzard, Avian/enzymology Kinetics Microscopy, Electron Muscle, Smooth/enzymology Myosins/metabolism,ultrastructure Phosphorus Radioisotopes Phosphorylation Turkeys
Chemicals
Actins Phosphorus Radioisotopes Myosins
Authors & Affiliations
1 authors, click to expand affiliations / ORCID
Trybus K M
Rosenstiel Basic Medical Sciences Research Center, Brandeis University, Waltham, Massachusetts 02254-9110.
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31 references, click to expand
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
Published
1989-12-00
Pages
2887-94
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2115938
Subset
IM
Grants
NIAMS NIH HHS · AR17350 · United States
NHLBI NIH HHS · HL38113 · United States
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