Photoaffmity cross‐linking of F1ATPase from the thermophilic bacterium PS 3 by 3'‐arylazido‐β‐alanyl‐8‐azido ATP

FEBS Letters

Kagawa, Y.²  Dose, Klaus¹  Rathgeber, Gabriele¹  Masafumi, Y.¹  Schäfer, Hans-Jochen¹ 

[1] Institut für Biochemie, Johannes Gutenberg-Universität, J.-J.-Becher-Weg 30, D-6500 Mainz, FRG;Department of Biochemistry, Jichi Medical School, Minamikawachi-machi, Tochigi-ken 329-04, Japan

关键词: Bacterial F1ATPase;    Thermophilic bacterium PS3;    Photoaffinity cross-linking;    Nucleotide-binding site α/ β interface;    TF1;    coupling factor 1 (F1ATPase) from the thermophilic bacterium PS3;    DiN3ATP;    3'-arylazido-β-alanyl-8-azido ATP;    3'-O-{3-[N-(4-azido-2-nitrophenyl)amino]propionyl}-8-azidoadenosine 5'-triphosphate;    8-N3ATP;    8-azido ATP;    8-azidoadenosine 5'-triphosphate;   

DOI  :  10.1016/0014-5793(85)80724-9

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

To study the localization of the nucleotide binding sites of coupling factor 1 (TF₁) from the thermophilic bacterium PS3 we used the bifunctional (cross-linking) 3'-arylazido-β-alanyl-8-azido ATP (DiN₃ATP) for photoaffinity labeling. DiN₃ATP is hydrolyzed by TF₁ in the absence of ultraviolet light. Irradiation (UV light) of TF₁ in the presence of DiN₃ATP results in a nucleotide-specific reduction of ATPase activity and in a nucleotide-specific formation of different cross-linked proteins (dimers, trimers, oligomers) formed by the major subunits α and/or β. The results suggest that nucleotide binding sites (one, two, possibly all) are located at the interfaces between these subunits.

【 授权许可】

Unknown