PMID: 2502175May 2, 1989Paper

Serine-324 of myosin's heavy chain is photoaffinity-labeled by 3'(2')-O-(4-benzoylbenzoyl)adenosine triphosphate

Biochemistry
R MahmoodR G Yount

Abstract

A portion of the active site of rabbit skeletal myosin near the ribose ring of ATP can be labeled by the photoaffinity analogue 3'(2')-O-(4-benzoylbenzoyl)adenosine triphosphate (Bz2ATP). The specificity of the photolabeling was assured by first trapping [14C]Bz2ATP at the active site by use of thiol cross-linking agents [Mahmood, R., Cremo, C., Nakamaye, K., & Yount, R. (1987) J. Biol. Chem. 262, 14479-14486]. Five radioactive peptides were isolated by high-performance liquid chromatography after extensive trypsin and subtilisin digestion of photolabeled myosin subfragment 1. Four of these peptides were sequenced by Edman techniques, and all originated from a region with the sequence Gly-Glu-Ile-Thr-Val-Pro-Ser-Ile-Asp-Asp-Gln, which corresponds to rabbit myosin heavy chain residues 318-328. The fifth labeled peptide had an amino acid composition appropriate for residues 312-328. Amino acid composition, radiochemical analysis, and sequence data indicate that Ser-324 is the major amino acid residue photolabeled by Bz2ATP. Spectrophotometric evidence indicates that the benzophenone carbonyl group has inserted into a C-H bond from either the alpha- or beta-carbon of serine. These results place Ser-324 at a distance of 6-7 A from ...Continue Reading

References

Oct 1, 1979·Proceedings of the National Academy of Sciences of the United States of America·J A Wells, R G Yount
Mar 1, 1985·Proceedings of the National Academy of Sciences of the United States of America·Y Okamoto, R G Yount
Dec 1, 1986·Proceedings of the National Academy of Sciences of the United States of America·H M WarrickJ A Spudich
May 16, 1973·Journal of the American Chemical Society·R BreslowW Washburn
Jul 1, 1983·Proceedings of the National Academy of Sciences of the United States of America·J KarnL Barnett
Jul 1, 1984·Proceedings of the National Academy of Sciences of the United States of America·I Rayment, D A Winkelmann
Jan 1, 1972·Methods in Enzymology·W R Gray

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Citations

Oct 19, 2000·Biophysical Chemistry·B JohnA K Lala
Sep 13, 2005·Journal of Molecular Biology·Becky M MillerSanford I Bernstein
Jul 1, 1995·Bioconjugate Chemistry·J D OlszewskiG D Prestwich
Sep 29, 2018·Science Advances·Jun-Hoe LeeMichael Hiller
May 17, 1994·Biochemistry·G Dormán, G D Prestwich
Aug 8, 1997·Journal of Molecular Biology·S I Bernstein, R A Milligan

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