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Chicken smooth muscle myosin light chain kinase is acetylated on its NH2-terminal methionine

  • Protein Kinases
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Abstract

The reported cDNA structrre, of chicken smooth muscle myosin light chain kinase (smMLCK) encodes a protein of 972 residues (Olsonet al. Proc. Natl. Acad. Sci USA, 87: 2284–2288, 1990). The calculated Mr is 107, 534 whereas the estimate by SDS-PAGE is approximately 130, 000. Gibson and Higgins (DNA Sequence (in press)) have recently reported the possibility of errors, in the cDNA sequence for non-muscle MLCK and that the NH2-terminus of both it and smMLCK may extend beyond the reported coding region. The native smMLCK is NH2-terminally blocked. A CNBr peptide derived from smMLCK contains the NH2-terminal sequence Asp-Phe-Arg-Ala corresponding to residues 2 to 4 in the smMLCK sequence indicating, that Met-1 is present. Using a limited thermolysin digest we isolated an NH2-terminally blocked peptide by reversed-phase HPLC. This thermolytic peptide had a mass of approximately 797 by time of flight mass spectrometry. Amino acid analysis and Edman sequencing of a CNBr-subfragment of the thermolytic peptide indicated that it had the composition and sequence, (Met)-Asp-Phe-Arg-Ala-Asn, with a calculated mass of 753. The difference in mass corresponds to the NH2-terminal Met being blocked by actylation. The results demonstrate that the NH2-terminal sequence of smMLCK inferred from the reported cDNA sequence is correct and that the proposed initiating, Met is not removed, but modified by α-NH2 acetylation of the translation product.

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Authors and Affiliations

  1. St Vincent's Institute of Medical Research, 41 Victoria Parade, 3065, Fitzroy, Vic, Australia

    Maree C. Faux, Kenneth I. Mitchelhill, Frosa Katsis, Richard E. H. Wettenhall & Bruce E. Kemp

  2. Department of Biochemistry, The University of Melbourne, 3052, Parkville, Vic, Australia

    Richard E. H. Wettenhall

Authors
  1. Maree C. Faux
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  2. Kenneth I. Mitchelhill
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  3. Frosa Katsis
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  4. Richard E. H. Wettenhall
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  5. Bruce E. Kemp
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Faux, M.C., Mitchelhill, K.I., Katsis, F. et al. Chicken smooth muscle myosin light chain kinase is acetylated on its NH2-terminal methionine. Mol Cell Biochem 127, 81–91 (1993). https://doi.org/10.1007/BF01076759

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