2,8-Diazido-ATP--a short-length bifunctional photoaffinity label for photoaffinity cross-linking of a stable F1 in ATP synthase (from thermophilic bacteria PS3).
To demonstrate the direct interfacial position of nucleotide binding sites between subunits of proteins we have synthesized the bifunctional photoaffinity label 2,8-diazidoadenosine 5'-triphosphate (2,8-DiN3ATP). UV irradiation of the F1-ATPase (TF1) from the thermophilic bacterium PS3 in the presence of 2,8-DiN3ATP results in a nucleotide-dependent inactivation of the enzyme and in a nucleotide-dependent formation of alpha-beta crosslinks. The results confirm an interfacial localization of all the nucleotide binding sites on TF1.
Institut für Biochemie, Johannes Gutenberg-Universität, Mainz, Germany.
H J
HJ
Schäfer
G
G
Rathgeber
Y
Y
Kagawa
D000255:Adenosine Triphosphate / Q000031:analogs & derivatives* / Q000737:chemistry; D000345:Affinity Labels / Q000737:chemistry*; D001386:Azides / Q000737:chemistry*; D001419:Bacteria / Q000201:enzymology*; D003432:Cross-Linking Reagents / Q000737:chemistry*; D015151:Immunoblotting; D008027:Light; D008958:Models, Molecular; D006180:Proton-Translocating ATPases / Q000737:chemistry* / Q000378:metabolism / Q000528:radiation effects
D000345:Affinity Labels; D001386:Azides; D003432:Cross-Linking Reagents; C097709:2,8-diazidoadenosine 5'-triphosphate; D000255:Adenosine Triphosphate; D006180:Proton-Translocating ATPases