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Mechanism of phosphorylation of the regulatory light chain of myosin from tarantula striated muscle

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Abstract

Contraction is modulated in many striated muscles by Ca2+-calmodulin dependent phosphorylation of the myosin regulatory light chain (RLC) by myosin light chain kinase. We have investigated the biochemical mechanism of RLC phosphorylation in tarantula muscle to better understand the basis of myosin-linked regulation. In an earlier study it was concluded that the RLC occurred as two species, both of which could be phosphorylated, potentiating contraction. Here we present evidence that only a single species exists, and that this can be phosphorylated at one or two sites. In relaxed muscle we find evidence for a substantial level of basal phosphorylation at the first site. This is augmented on activation, followed by partial phosphorylation of the second site. We find in addition that Ca2+ has a dual effect on light chain phosphorylation, depending on its concentration. At low concentration (relaxing conditions) only basal phosphorylation is observed, while at higher concentrations (activating conditions) RLC phosphorylation is stimulated. At still higher Ca2+ concentrations we find partial inhibition of RLC phosphorylation, suggesting an additional mechanism by which the muscle cell can fine tune contractile activity by controlling the level of free Ca2+.

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Authors and Affiliations

  1. Departamento de Biología Estructural, Instituto Venezolano de Investigaciones Científicas (IVIC), Apdo. 21827, Caracas, 1020A, Venezuela

    Carlos Hidalgo & Raúl Padrón

  2. Department of Cell Biology, University of Massachusetts Medical School, 55 Lake Avenue North, Worcester, Massachusetts, 01655, USA

    Carlos Hidalgo & Roger Craig

  3. Department of Physiology, University of Massachusetts Medical School, 55 Lake Avenue North, Worcester, Massachusetts, 01655, USA

    Mitsuo Ikebe

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  1. Carlos Hidalgo
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  2. Roger Craig
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  3. Mitsuo Ikebe
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  4. Raúl Padrón
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Hidalgo, C., Craig, R., Ikebe, M. et al. Mechanism of phosphorylation of the regulatory light chain of myosin from tarantula striated muscle. J Muscle Res Cell Motil 22, 51–59 (2001). https://doi.org/10.1023/A:1010388103354

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