Amino acid sequences of myosin essential and regulatory light chains from two clam species: comparison with other molluscan myosin light chains.
We have determined the amino acid sequences of the essential light chains (ELC) and regulatory light chains (RLC) of myosin from two species of clam, Mercenaria mercenaria and Macrocallista nimbosa, using protein chemistry methods. The N-termini of all four proteins were blocked, and sequencing was carried out on various chemically and enzymatically produced peptide fragments. Cleavage of either Mercenaria RLC (MRLC) or Macrocallista RLC (VLC) at its 3 Arg yielded four peptides, three of which could not be sequenced directly, due to an N-terminal blocking group and 2 Arg-Gln bonds in these proteins. The fourth peptide was partially and specifically cleaved at an unusually reactive residue, Met-64, which is invariant in all known RLC sequences. A comparison of all available molluscan ELC and RLC sequences was carried out in search of clues to functionally important features of these proteins in muscles which are regulated by a Ca(2+)-sensitive myosin. By analogy with other RLCs, VRLC and MRLC may be phosphorylated at Ser-11 by an endogenous kinase. All myosin light chains, like troponin C and calmodulin, contain four homologous regions, I to IV, each of which contains a twelve-residue potential Ca(2+)-binding loop flanked on either side by a pair of helices. All RLCs, including those from Ca(2+)-insensitive myosins, contain a divalent cation-binding site in region I. Clam and other molluscan ELCs contain a single Ca(2+)-binding site in region III. This site is present only in the ELCs of myosins that are regulated by direct binding of Ca2+. The ELC site III is likely to play a key role in the regulation of molluscan muscle contraction.
Barouch
W W
WW
Department of Biological Chemistry, School of Medicine, University of Maryland, Baltimore 21201.
Breese
K E
KE
Davidoff
S A
SA
Leszyk
J
J
Szent-Györgyi
A G
AG
Theibert
J L
JL
Collins
J H
JH
D000595:Amino Acid Sequence; D000818:Animals; D049872:Bivalvia / Q000032:analysis*; D002118:Calcium / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D008969:Molecular Sequence Data; D009218:Myosins / Q000737:chemistry* / Q000378:metabolism; D010766:Phosphorylation
D009218:Myosins; D002118:Calcium