Direct Sequence Analysis of a Cyanogen Bromide-Generated Peptide Corresponding to the Region of Subfragment-1 of Adult Chicken Gizzard Myosin Heavy Chain Predicted to Contain a Seven-Amino Acid Insertion

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The amino acid sequence of a cyanogen bromide-generated peptide corresponding to the region of subfragment-1 of chicken gizzard myosin, predicted from the cDNA sequence to contain a seven-amino acid insertion, was determined directly. Subfragment-1 was prepared from adult chicken gizzard myosin by limited digestion with papain, and a 137-residue peptide was obtained by cleaving subfragment-1 with cyanogen bromide. Amino acid composition analysis and sequence analysis of the fragments generated from this peptide by formic acid or a-chymotrypsin revealed that the peptide corresponded to leucine-165 to methionine-301 of chicken gizzard myosin heavy chain previously deduced from the cDNA sequence by Yanagisawa, M. et al.. The predicted seven-amino acid insertion (Gln-Gly-Pro-Ser-Phe-Ser-Tyr), absent from other smooth muscle myosin,was thus confirmed to be present in chicken gizzard myosin. The amino acid sequence surrounding the insertion site of chicken gizzard myosin shows greater homology to the corresponding sequence of other known smooth muscle and non-muscle myosins than to those of chicken skeletal and cardiac muscle myosins. The insertion sequence may confer gizzard-specific functions to the gizzard myosin heavy chain, whereas the amino acid sequence surrounding the insertion site may be specific to smooth muscle.
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