Comparative structure of the protease-sensitive regions of the subfragment-1 heavy chain from smooth and skeletal myosins.
The heavy chain fragments generated by restricted proteolysis of the smooth chicken gizzard myosin subfragment-1 (S-1) with trypsin, Staphylococcus aureus V8 protease, and chymotrypsin were isolated and submitted to partial amino acid sequencing. The comparison between the smooth and striated muscle myosin sequences permitted the unambiguous structural characterization of the two protease-vulnerable segments joining the three putative domain-like regions of the smooth head heavy chain. The smooth carboxyl-terminal connector is a serine-rich region located around positions 632-640 of the rabbit skeletal sequence and would represent the "A" site that is conformationally sensitive to the myosin 10 S-6 transition and to its interaction with actin (Ikebe, M., and Hartshorne, D. J. (1986) Biochemistry 25, 6177-6185). A third site which undergoes a nucleotide-dependent chymotryptic cleavage which inactivates the Mg2+-ATPase (Okamoto, Y., and Sekine, T. (1981) J. Biochem. (Tokyo) 90, 833-842, 843-849) was identified at Trp-31/Ser-32. It is vicinal to Lys-34 that is monomethylated in the skeletal heavy chain but not at all in the smooth sequence. However, the two trimethyl lysine residues present in the skeletal sequence are conserved in the same regions of the smooth S-1 and may play a general functional role in myosin. The smooth central 50-kDa segment could be selectively destroyed by a mild tryptic digestion in the absence of any unfolding agent, with a concomitant inhibition of the ATPase activities. This feature is in line with the proposed domain structure of the S-1 heavy chain and also suggests a relationship between the specific biochemical properties of the smooth S-1 and the particular conformation of its 50-kDa region.
Bonet
A
A
Centre de Recherche de Biochimie Macromoléculaire, Université de Montpellier I, France.
Mornet
D
D
Audemard
E
E
Derancourt
J
J
Bertrand
R
R
Kassab
R
R
D000595:Amino Acid Sequence; D000818:Animals; D002645:Chickens; D005895:Gizzard, Avian / Q000201:enzymology; D008969:Molecular Sequence Data; D009130:Muscle, Smooth / Q000201:enzymology*; D009132:Muscles / Q000201:enzymology*; D015879:Myosin Subfragments; D009218:Myosins / Q000032:analysis*; D010446:Peptide Fragments / Q000032:analysis*; D010447:Peptide Hydrolases / Q000378:metabolism*; D010449:Peptide Mapping; D011817:Rabbits
D015879:Myosin Subfragments; D010446:Peptide Fragments; D010447:Peptide Hydrolases; D009218:Myosins