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Molecular Dissection of Regulatory Light Chain Function in Vertebrate Smooth Muscle Myosins

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Regulation of the Contractile Cycle in Smooth Muscle

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Summary

The recent elucidation of the crystal structures of the myosin head and regulatory domain allows us to analyze the changes in myosin structure that occur during regulation. The light chains are the regulatory subunits and are bound to the α-helical segment of the heavy chain (regulatory domain), which extends from the motor domain (containing the ATPase and actin-binding sites) to the head-tail junction. We have used a recombinant DNA approach to investigate how the light chains in this location regulate the motor domain. To identify the subdomains/regions important for regulatory function, we have synthesized a series of mutant regulatory light chains (RLCs) and characterized them by their ability to restore Ca2+ regulation to the desensitized scallop myosin test system. Assays on chimeric RLCs composed of subdomains derived from vertebrate smooth muscle and skeletal muscle myosin RLCs demonstrate that the origin of the third subdomain specifies the regulatory capability of the RLC. A series of smooth muscle myosin RLC mutants with deletions in the fourth subdomain show that the C-terminal helix in this subdomain is essential for regulation. Although the skeletal and scallop RLCs need an intact Ca2+/Mg2+-binding site in the first subdomain for function, the smooth muscle RLC behaves differently and does not require divalent metal binding to this site for regulation. These studies demonstrate the regions of the RLC that are important for regulatory function.

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Author information

Authors and Affiliations

  1. MRC Laboratory of Molecular Biology, Hills Road, Cambridge, CB2 2QH, UK

    John Kendrick-Jones, Tony Rowe & Ana C. Rasera da Silva

  2. Department of Biochemistry, Instituto de Química, Universidade de São Paulo, CP 20780, CEP 01498, São Paulo, SP, Brazil

    Fernando C. Reinach

Authors
  1. John Kendrick-Jones
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  2. Tony Rowe
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  3. Ana C. Rasera da Silva
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  4. Fernando C. Reinach
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Editors and Affiliations

  1. Mie University of School Medicine, 2-175 Edobashi, Tsu, Mie, 514, Japan

    Takeshi Nakano M.D., Ph.D. (Professor) (Professor)

  2. University of Arizona, 85721, Tucson, AZ, USA

    David J. Hartshorne Ph.D. (Professor) (Professor)

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© 1995 Springer-Verlag Tokyo

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Kendrick-Jones, J., Rowe, T., Rasera da Silva, A.C., Reinach, F.C. (1995). Molecular Dissection of Regulatory Light Chain Function in Vertebrate Smooth Muscle Myosins. In: Nakano, T., Hartshorne, D.J. (eds) Regulation of the Contractile Cycle in Smooth Muscle. Springer, Tokyo. https://doi.org/10.1007/978-4-431-65880-1_7

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  • DOI: https://doi.org/10.1007/978-4-431-65880-1_7

  • Publisher Name: Springer, Tokyo

  • Print ISBN: 978-4-431-65882-5

  • Online ISBN: 978-4-431-65880-1

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