Selective removal of the carboxyl-terminal tail end of the Dictyostelium myosin II heavy chain by chymotrypsin.
Dictyostelium myosin II is a conventional myosin consisting of two heavy chains of 243,000 Da and two pairs of light chains of 16,000 and 18,000 Da. In this paper, we show that the heavy chain of myosin II can be rapidly and selectively cleaved by chymotrypsin to yield two fragments with molecular weights of 195,000 and 38,000 Da as estimated from sodium dodecyl sulfate-polyacrylamide gels. Chymotryptic cleavage at this site occurs most readily in the absence of salt and is greatly inhibited as the salt concentration is increased from 0 to 60 mM. Amino acid sequence analysis of the small fragment demonstrates that its amino terminus corresponds to lysine 1826 of the myosin II heavy chain. If the fragment extends to the carboxyl terminus of the myosin II heavy chain, it would have a molecular weight of 33,700. Upon digestion of myosin II which has been phosphorylated with a high molecular weight Dictyostelium myosin heavy chain kinase (Côté, G.P., and Bukiejko, U. (1987) J. Biol. Chem. 262, 1065-1072), all of the phosphate is recovered on the 33,700-Da tail-end fragment. Chymotrypsin-cleaved myosin II is shown to be capable of forming filaments at salt concentrations between 20 and 100 mM as judged by its ability to be sedimented by centrifugation. Only the large fragment of myosin II is found in the pellet; the 33,700-dalton fragment remains soluble. Chymotrypsin-cleaved myosin II can bind to actin and displays a high Ca2+-activated ATPase activity but has very low actin-activated ATPase activity.
D000199:Actins / Q000378:metabolism; D000251:Adenosine Triphosphatases / Q000378:metabolism; D000595:Amino Acid Sequence; D002918:Chymotrypsin / Q000378:metabolism*; D004023:Dictyostelium / Q000378:metabolism*; D007700:Kinetics; D008970:Molecular Weight; D015879:Myosin Subfragments; D009218:Myosins / Q000378:metabolism; D009994:Osmolar Concentration; D010446:Peptide Fragments / Q000032:analysis / Q000378:metabolism; D010766:Phosphorylation; D012995:Solubility
D000199:Actins; D015879:Myosin Subfragments; D010446:Peptide Fragments; D002918:Chymotrypsin; D000251:Adenosine Triphosphatases; D009218:Myosins