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In vitro production of enzymatically active myosin heavy chain

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Summary

In order to initiate studies on the structural and functional relationships of the myosin heavy chain, we constructed a full-length complementary DNA encoding the isoform that is found in the fast white muscle of the embryonic chicken. The complementary DNA contained 108 basepairs of its 3′-untranslated region and was preceded by a leader sequence derived from the alfalfa mosaic virus. Similarly, a complementary DNA encoding 963 amino acids which encompass the subfragment-1 of myosin and part of the subfragment-2 was also constructed. Each was inserted into the expression vector pMT2 and transiently transfected into COS-1 cells. Both constructs directed the expression of the respective proteins, each of which was immunogenic. The full-length and subfragment-1 proteins interacted with actin and demonstrated high levels of a K+-actived, EDTA-resistant ATPase activity, which is characteristic of myosin.

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Authors and Affiliations

  1. Department of Pharmacology and Cell Biophysics, University of Cincinnati, College of Medicine, 231 Bethesda Avenue, 45267, Cincinnati, OH, USA

    H. Rindt & J. Robbins

  2. Department of Internal Medicine, University of Cincinnati, College of Medicine, 231 Bethesda Avenue, 45267, Cincinnati, OH, USA

    B. J. Bauer

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  1. H. Rindt
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  2. B. J. Bauer
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  3. J. Robbins
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Rindt, H., Bauer, B.J. & Robbins, J. In vitro production of enzymatically active myosin heavy chain. J Muscle Res Cell Motil 14, 26–34 (1993). https://doi.org/10.1007/BF00132177

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  • DOI: https://doi.org/10.1007/BF00132177

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