A short hydrophobic segment next to tryptophan-130 in myosin heavy chain is close to the ribose ring of ADP bound in the adenosinetriphosphatase site.
The ATPase site of rabbit skeletal myosin was covalently labeled by an ADP analogue that carried a biotin moiety on its adenine ring and a photoreactive phenyl azide group on its ribose ring [Sutoh, K., Yamamoto, K., & Wakabayashi, T. (1986) Proc. Natl. Acad. Sci. U.S.A. 83, 212-216]. The ADP analogue was tightly trapped into the ATPase site in the presence of vanadate ions and then covalently cross-linked to the site by UV irradiation. The N-terminal 23,000-dalton tryptic fragment of the heavy chain was selectively labeled with the analogue. Further mapping of the labeled segment along the 23-kDa fragment was carried out by "end-label fingerprinting" which employed site-directed antibodies against both ends of the N-terminal heavy chain fragment. The mapping revealed that a hydrophobic segment of approximately 10 residues next to Trp-130, which was reported to be in proximity to the adenine ring of ADP bound to the ATPase site [Okamoto, Y., & Yount, R. G. (1985) Proc. Natl. Acad. Sci. U.S.A. 82, 1575-1579], was the site of covalent labeling with the ADP analogue. The result indicates that the hydrophobic segment is close to the ribose ring of ADP bound to the ATPase site.
D000251:Adenosine Triphosphatases / Q000378:metabolism*; D000818:Animals; D000906:Antibodies; D000936:Antigen-Antibody Complex; D001665:Binding Sites; D001710:Biotin; D008970:Molecular Weight; D009132:Muscles / Q000201:enzymology; D015879:Myosin Subfragments; D009218:Myosins / Q000378:metabolism*; D010446:Peptide Fragments / Q000378:metabolism*; D010449:Peptide Mapping; D011817:Rabbits; D014364:Tryptophan
D000906:Antibodies; D000936:Antigen-Antibody Complex; D015879:Myosin Subfragments; D010446:Peptide Fragments; D001710:Biotin; D014364:Tryptophan; D000251:Adenosine Triphosphatases; D009218:Myosins