Photoaffinity labeling of the coupling factor 1 from the thermophilic bacterum PS3 by 8‐azido ATP

FEBS Letters

Dose, Klaus²  Rathgeber, Gabriele²  Scheurich, Peter²  Kagawa, Yasuo¹  Schäfer, Hans-Jochen² 

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

关键词: Bacterial F1ATPase;    Thermophilic bacterium PS3;    Photoaffinity labeling;    Catalytic nucleotide binding site;    Noncatalytic nucleotide binding site;    TF1;    coupling factor 1 (F1ATPase) from the thermophilic bacterium PS3;    8-N3ATP;    8-azido ATP;    8-azidoadenosine 5'-triphosphate;    3'-arylazido-β-alanyl-ATP;    3'-O-{3-[N-(4-azido-2-nitrophenyl)amino]propionyl} adenosine 5'-triphosphate;    3'-arylazido-β-alanyl-8-azido ATP;    3'-O-{3-[N-(4-azido-2-nitro- phenyl)amino]propionyl}-8-azidoadenosine 5'-triphosphate;    8-N3AMP;    8-azidoadenosine 5'-monophosphate;    2-N3ATP;    2-azidoadenosine 5'-triphosphate;   

DOI  :  10.1016/0014-5793(84)81079-0

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

To localize the nucleotide binding sites of the F₁ATPase (TF₁) from the thermophilic bacterium PS3 we have used ¹⁴C-labeled 8-azido ATP (8-N₃ATP) as photoaffmity label. 8-N₃ATP is hydrolyzed by the F,ATPase in the absence of ultraviolet light. Irradiation by ultraviolet light of the enzyme in the presence of 8-N₃ATP results in reduction of ATPase activity and in preferential nucleotide specific labeling of the α subunits (0.8–0.9 mol 8-N₃ATP/TF₁,α:β = 4:1). Inactivation and labeling do not depend on the presence of Mg²⁺. Both effects decrease upon addition of various nucleotide di- or triphosphates.

【 授权许可】

Unknown