Characterization of nucleotide‐binding sites on the chloroplast coupling factor 1 using two photolabile analogs

FEBS Letters

Shavit, Noun¹  Selman, Bruce R.²  Selman-Reimer, Susanne²  Abbott, Marilyn S.² 

[1] Department of Biology, Ben-Gurion University of the Negev, 84105 Beer Sheva, Israel;Department of Biochemistry, College of Agricultural and Life Sciences, University of Wisconsin-Madison, Madison, WI 53706, USA

关键词: Coupling factor;    ATPase;    Photoaffinity analog;    Chloroplast;    Azido-ADP 2';    3'-O-(4-Benzoyl)benzoyl-ADP;    CF1;    chloroplast coupling factor 1;    2-N3-ADP or 2-N3-ATP;    2-azidoadenosine 5'-diphosphate or -triphosphate;    Bz-ADP or Bz-ATP;    2';    3'-O-(4-benzoyl)benzoyladenosine 5'-diphosphate or -triphosphate;    octylglucoside;    n-octyl β-D-glucopyranoside;    LDS;    lithium dodecyl sulfate;    chl;    chlorophyll;    CCCP;    carbonyl cyanide m-chlorophenylhydrazone;    DTT;    dithiothreitol;   

DOI  :  10.1016/0014-5793(86)81102-4

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Nucleotide-binding sites on the chloroplast coupling factor 1 (CF₁) have been probed using two photoreactive ADP analogs: 2-azido-ADP (2-N₃-ADP) and 2',3'-O-(4-benzoyl)benzoyl-ADP (Bz-ADP). Photolabeling of the isolated CF₁ with 2-N₃-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 CF₁ 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 CF₁. Bz-ADP, however, labels the binding site(s) on membrane-bound CF₁ in a slightly different manner.

【 授权许可】

Unknown