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Photoaffinity labeling of the coupling factor 1 from the thermophilic bacterum PS3 by 8‐azido ATP
Author(s) -
Schäfer Hans-Jochen,
Scheurich Peter,
Rathgeber Gabriele,
Dose Klaus,
Kagawa Yasuo
Publication year - 1984
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(84)81079-0
Subject(s) - nucleotide , atpase , thermophile , atp hydrolysis , chemistry , enzyme , ultraviolet light , biochemistry , photoaffinity labeling , adenosine triphosphate , binding site , photochemistry , gene
To localize the nucleotide binding sites of the F 1 ATPase (TF 1 ) from the thermophilic bacterium PS3 we have used 14 C‐labeled 8‐azido ATP (8‐N 3 ATP) as photoaffmity label. 8‐N 3 ATP is hydrolyzed by the F,ATPase in the absence of ultraviolet light. Irradiation by ultraviolet light of the enzyme in the presence of 8‐N 3 ATP results in reduction of ATPase activity and in preferential nucleotide specific labeling of the α subunits (0.8–0.9 mol 8‐N 3 ATP/TF 1 ,α:β = 4:1). Inactivation and labeling do not depend on the presence of Mg 2+ . Both effects decrease upon addition of various nucleotide di‐ or triphosphates.