Open AccessSynthesis of a New Reagent, Ethyl 4‐azidobenzoylaminoacetimidate, and Its Use for RNA‐Protein Cross‐linking within Escherichia coli Ribosomal 30‐S Subunits
Author(s) -
MILLON Régine,
OLOMUCKI Martin,
GALL JeanYves,
GOLINSKA Barbara,
EBEL JeanPierre,
EHRESMANN Bernard
Publication year - 1980
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1980.tb04890.x
Subject(s) - reagent , escherichia coli , rna , ribosomal protein , ribosomal rna , chemistry , oligonucleotide , biochemistry , microbiology and biotechnology , protein subunit , 50s , polyacrylamide gel electrophoresis , biology , ribosome , gene , enzyme , organic chemistry
A new reagent, ethyl 4‐azidobenzoylaminoacetimidate, was prepared in a four‐step synthesis starting from 4‐aminobenzoic acid. This compound was used to cross‐link RNA with proteins within the Escherichia coli 30‐S ribosomal subunits. Following the reaction of the imidoester function with protein NH 2 groups, photoactivation of the azide binds the other end of the reagent to RNA. The cross‐linked proteins were labelled with 125 I and identified by bidimensional gel electrophoresis. Proteins S3, S4, SS, S7, S9, S17, S18 and, in a lower and more variable yield, S12, S13, S14 and S16 were bound to 16‐S RNA. These results were confirmed by isolating cross‐linked protein‐oligonucleotide complexes from 30‐S subunits containing 32 P‐labelled RNA.