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Characterization of nucleotide‐binding sites on the chloroplast coupling factor 1 using two photolabile analogs
Author(s) -
Abbott Marilyn S.,
Shavit Noun,
Selman-Reimer Susanne,
Selman Bruce R.
Publication year - 1986
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(86)81102-4
Subject(s) - nucleotide , binding site , chemistry , stereochemistry , chloroplast , protein subunit , biochemistry , gene
Nucleotide‐binding sites on the chloroplast coupling factor 1 (CF 1 ) have been probed using two photoreactive ADP analogs: 2‐azido‐ADP (2‐N 3 ‐ADP) and 2',3'‐ O ‐(4‐benzoyl)benzoyl‐ADP (Bz‐ADP). Photolabeling of the isolated CF 1 with 2‐N 3 ‐ADP results in incorporation of the analog exclusively into the β‐subunit of the enzyme. The location of the nucleotide‐binding site(s) within the β‐subunit of the CF 1 was investigated using peptide mapping. Within the discrimination limits of this technique, it is concluded that the azidoand benzoyl‐modified analogs both bind to the same conformation of the nucleotide‐binding site(s) of soluble CF 1 . Bz‐ADP, however, labels the binding site(s) on membrane‐bound CF 1 in a slightly different manner.