caa a22 4500 475745337 CHVBK 20180406123511.0 cr unu---uuuuu 170329e20000701xx s 000 0 eng 10.1023/A:1005698407859 doi (NATIONALLICENCE)springer-10.1023/A:1005698407859 Primary structure of myosin from the striated adductor muscle of the Atlantic scallop, Pecten maximus, and expression of the Regulatory Domain [Elektronische Daten] [Daniel Janes, Hitesh Patel, Peter Chantler] We have determined the complete cDNA and deduced amino acid sequences of the heavy chain, regulatory light chain and essential light chain which constitute the molecular structure of myosin from the striated adductor muscle of the scallop, Pecten maximus. The deduced amino acid sequences of P. maximus regulatory light chain, essential light chain and heavy chain comprise 156, 156 and 1940 amino acids, respectively. These myosin peptide sequences, obtained from the most common of the eastern Atlantic scallops, are compared with those from three other molluscan myosins: the striated adductor muscles of Argopecten irradians and Placopecten magellanicus, and myosin from the siphon retractor muscle of the squid, Loligo pealei. The Pecten heavy chain sequence resembles those of the other two scallop sequences to a much greater extent as compared with the squid sequence, amino acid identities being 97.5% (A. irradians), 95.6% (P. magellanicus) and 73.6% (L. pealei), respectively. Myosin heavy chain residues that are known to be important for regulation are conserved in Pecten maximus. Using these Pecten sequences, we have overexpressed the regulatory light chain, and a combination of essential light chain and myosin heavy chain fragment, separately, in E. coli BL21 (DE3) prior to recombination, thereby producing Pecten regulatory domains without recourse to proteolytic digestion. The expressed regulatory domain was shown to undergo a calcium-dependent increase (∼7%) in intrinsic tryptophan fluorescence with a mid-point at a pCa of 6.6. Kluwer Academic Publishers, 2000 Janes Daniel Unit of Molecular and Cellular Biology, Royal Veterinary College, University of London, Royal College Street, NW1 0TU, London, UK aut Patel Hitesh Unit of Molecular and Cellular Biology, Royal Veterinary College, University of London, Royal College Street, NW1 0TU, London, UK aut Chantler Peter Unit of Molecular and Cellular Biology, Royal Veterinary College, University of London, Royal College Street, NW1 0TU, London, UK aut Journal of Muscle Research & Cell Motility Kluwer Academic Publishers 21/5(2000-07-01), 415-422 0142-4319 21:5<415 2000 21 10974 https://doi.org/10.1023/A:1005698407859 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1023/A:1005698407859 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Janes Daniel Unit of Molecular and Cellular Biology, Royal Veterinary College, University of London, Royal College Street, NW1 0TU, London, UK aut NATIONALLICENCE 700 1- Patel Hitesh Unit of Molecular and Cellular Biology, Royal Veterinary College, University of London, Royal College Street, NW1 0TU, London, UK aut NATIONALLICENCE 700 1- Chantler Peter Unit of Molecular and Cellular Biology, Royal Veterinary College, University of London, Royal College Street, NW1 0TU, London, UK aut NATIONALLICENCE 773 0- Journal of Muscle Research & Cell Motility Kluwer Academic Publishers 21/5(2000-07-01), 415-422 0142-4319 21:5<415 2000 21 10974 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer