A new simpler photoaffinity analogue of peptidyl tRNA.
The synthesis of the n-hydroxysuccinimide ester of N-(2-nitro-4-azidophenyl)glycine (NAG) is described. This reacts with E. coli phe-tRNA(Phe) to yield the photoaffinity label NAG-Phe-tRNA(Phe). This peptidyl tRNA analogue binds correctly to the peptidyl site of the E. coli ribosome. The only significant covalent products found after irradiation of a peptidyl site bound NAG-Phe-tRNA(Phe)-70S-poly(U) complex are 50S proteins L11 and L18. After irradiation the complex can still bind [(3)H]Phe-tRNA to the amino acyl site and participate in peptide bond formation with the covalently attached NAG-Phe moiety. Alternatively, one can allow peptide bond formation to occur first, prior to photolysis. The reaction products are still L11 and L18. Hence, both of these two proteins appear to be centrally located at the peptidyl transferase center.
D000217:Acyltransferases / Q000378:metabolism; D001386:Azides / Q000138:chemical synthesis; D001665:Binding Sites; D004926:Escherichia coli / Q000378:metabolism; D013058:Mass Spectrometry; D009596:Nitrophenols; D010455:Peptides; D010649:Phenylalanine; D011485:Protein Binding; D012343:RNA, Transfer / Q000031:analogs & derivatives* / Q000378:metabolism; D012270:Ribosomes / Q000378:metabolism
D001386:Azides; D009596:Nitrophenols; D010455:Peptides; D010649:Phenylalanine; D012343:RNA, Transfer; D000217:Acyltransferases