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)

摘要

>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 α-β crosslinks. The results confirm an interfacial localization of all the nucleotide binding sites on TF₁.