Serine-324 of myosin's heavy chain is photoaffinity-labeled by 3'(2')-O-(4-benzoylbenzoyl)adenosine triphosphate
- 1 May 1989
- journal article
- research article
- Published by American Chemical Society (ACS) in Biochemistry
- Vol. 28 (9) , 3989-3995
- https://doi.org/10.1021/bi00435a054
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., and 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 .ANG. from the 3''(2'') ribose oxygens of ATP bound at the active site of myosin.Keywords
This publication has 15 references indexed in Scilit:
- Conserved protein domains in a myosin heavy chain gene from Dictyostelium discoideum.Proceedings of the National Academy of Sciences, 1986
- Complete nucleotide and encoded amino acid sequence of a mammalian myosin heavy chain geneJournal of Molecular Biology, 1986
- 2-[(4-Azido-2-nitrophenyl)amino]ethyl triphosphate, a novel chromophoric and photoaffinity analog of ATP. Synthesis, characterization, and interaction with myosin subfragment 1Biochemistry, 1985
- Identification of an active site peptide of skeletal myosin after photoaffinity labeling with N-(4-azido-2-nitrophenyl)-2-aminoethyl diphosphate.Proceedings of the National Academy of Sciences, 1985
- Crystallization of myosin subfragment 1.Proceedings of the National Academy of Sciences, 1984
- Protein structural domains in the Caenorhabditis elegans unc-54 myosin heavy chain gene are not separated by introns.Proceedings of the National Academy of Sciences, 1983
- Exploring the adenine nucleotide binding sites on mitochondrial F1-ATPase with a new photoaffinity probe, 3'-O-(4-benzoyl)benzoyl adenosine 5'-triphosphate.Journal of Biological Chemistry, 1982
- Analysis of phenylthiohydantoin amino acids by high-performance liquid chromatography on DuPont Zorbax cyanopropylsilane columnsAnalytical Biochemistry, 1979
- Active site trapping of nucleotides by crosslinking two sulfhydryls in myosin subfragment 1.Proceedings of the National Academy of Sciences, 1979
- [8] End-group analysis using dansyl chloridePublished by Elsevier ,1972