Synthesis and application of bidentate photoaffinity cross-linking reagents. Nucleotide photoaffinity probes with two photoactive groups.

摘要: Two "targeted bidentate" photoaffinity cross-linking reagents, the monoanhydride of 8-N3ADP with N-(4-(benzoyl)phenylmethyl)phosphoramide ([gamma-32P]8-N3ATP gamma BP) and 8-N3GDP N-(4-(benzoyl)phenylmethyl)-phosphoramide ([gamma-32P]8-N3GTP BP), were developed for studying inter- intramolecular interactions nucleotide-binding proteins. Experiments using these bidentate reagents two photoactive groups led to specific cross-linking: [gamma-32P]8-N3GTP BP [gamma-32P]8-N3ATP showed intersubunit glutamate dehydrogenase appeared cross-link alpha- beta-subunits tubulin. The non-azido "monodentate" versions ADP ([gamma-32P]ATP GDP ([gamma-32P]GTP also synthesized characterized. ability monodentate one group serve as probes was established by photolabeling specifically exchangeable GTP-binding domain tubulin [gamma-32P]GTP ATP-binding purified adenylate kinase several proteins in human brain homogenate [gamma-32P]ATP BP.