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

Journal Article · · Biochemistry; (USA)
DOI:https://doi.org/10.1021/bi00435a054· OSTI ID:5434862
; ;  [1]
  1. Washington State Univ., Pullman (USA)
A portion of the active site of rabbit skeletal myosin near the ribose ring of ATP can be labeled by the photoaffinity analogue 3{prime}(2{prime})-O-(4-benzoylbenzoyl)adenosine triphosphate (Bz{sub 2}ATP). The specificity of the photolabeling was assured by first trapping ({sup 14}C)Bz{sub 2}ATP at the active site by use of thiol cross-linking agents. 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 312-328. 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 Bz{sub 2}ATP. 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 {angstrom} from the 3{prime}(2{prime}) ribose oxygens of ATP bound at the active site of myosin.
OSTI ID:
5434862
Journal Information:
Biochemistry; (USA), Journal Name: Biochemistry; (USA) Vol. 28:9; ISSN 0006-2960; ISSN BICHA
Country of Publication:
United States
Language:
English

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