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

Abstract
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.

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