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Cloning of the cDNA encoding human nonmuscle myosin heavy chain-B and analysis of human tissues with isoform-specific antibodies

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Summary

Previously, we reported the sequence of cDNA clones encoding amino acids 63 through 723 of the human nonmuscle myosin heavy chain-B isoform. In this paper, we present the derived sequence of the remaining 1303 amino acids along with 5′ and 3′ untranslated sequences. We made use of the differences between the derived nonmuscle myosin heavy chain-A and-B amino acid sequences to raise isoform-specific antibodies. Immunoblot analysis reveals a differential expression of both myosin heavy chain isoforms in a variety of human adult and foetal tissues and cells. When extracts of human adult aorta were subjected to gel electrophoresis, two distinct Coomassie Blue-stained bands and a fused band were seen migrating at approximately 200 kDa. These bands can be detected with four different specific antibodies recognizing the two different smooth muscle myosin heavy chain isoforms (204 kDa and 200 kDa) and the two different nonmuscle myosin heavy chain isoforms (A and B). Using immunohistochemistry, we confirmed the presence of the four different isoforms in adult and foetal aortas.

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Authors and Affiliations

  1. Laboratory of Molecular Cardiology National Heart, Lung, and Blood Institute, National Institutes of Health, 20892, Bethesda, MD, USA

    Carrie L. Phillips & Robert S. Adelstein

  2. Second Department of Internal Medicine, Yamaguchi University School of Medicine, 1144 Kogushi, 755, Ube, Yamaguchi Pref., Japan

    Katsutoshi Yamakawa

Authors
  1. Carrie L. Phillips
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  2. Katsutoshi Yamakawa
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  3. Robert S. Adelstein
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Present address: Laboratory of Pathology, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA.

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Phillips, C.L., Yamakawa, K. & Adelstein, R.S. Cloning of the cDNA encoding human nonmuscle myosin heavy chain-B and analysis of human tissues with isoform-specific antibodies. J Muscle Res Cell Motil 16, 379–389 (1995). https://doi.org/10.1007/BF00114503

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  • DOI: https://doi.org/10.1007/BF00114503

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