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PMID: 3540934 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Nucleotide binding by a 24-residue peptide from the RecA protein of Escherichia coli.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 83 ·No. 24 ·1986-12-00 ·Pages 9289-93

Knight KL, McEntee K

Abstract

We have recently demonstrated that two ATP analog affinity labels, 8-azidoadenosine 5'-triphosphate (N3ATP) and 5'-p-fluorosulfonylbenzoyladenosine (5'FSBA), covalently modify RecA protein of Escherichia coli at a specific tyrosine residue (Tyr-264) located within a 24-residue tryptic peptide (T-31) spanning residues 257-280. Here we show that N3ATP efficiently modifies purified peptide T-31 and show that the interaction is specific by the following criteria: photolabeling of peptide T-31 is saturable with respect to the N3ATP concentration; photolabeling is competitive with ATP and adenosine but not with adenine, UTP, or TTP; and other peptides derived from RecA protein were poor substrates for photolabeling except for one fragment that showed a nonspecific interaction with the photoaffinity analog. Analysis of N3ATP-modified T-31 shows that the photolabel attaches to more than one site within the peptide. These data argue that peptide T-31 contains some sites of contact for adenine and ribose moieties of ATP when it is bound to RecA protein.

MeSH Terms
Adenosine Triphosphate/analogs & derivatives,metabolism Affinity Labels Amino Acid Sequence Binding Sites Escherichia coli Peptide Fragments/metabolism Photochemistry Protein Binding Rec A Recombinases/metabolism Structure-Activity Relationship
Chemicals
Affinity Labels Peptide Fragments Adenosine Triphosphate Rec A Recombinases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Knight K L
McEntee K
References (35)
35 references, click to expand
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.83.24.9289
Published
1986-12-00
Pages
9289-93
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC387123
Subset
IM
Grants
NIGMS NIH HHS · GM29558 · United States
External Links
PubMed source DOI: 10.1073/pnas.83.24.9289 PMC Full Text View journal details
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