title: Stimulation, by two Escherichia coli supernatant proteins, of the initiation of polypeptide synthesis.
abstract: Two protein factors (A and B) have been partially purified from Escherichia coli supernatant which, in combination, are more effective than 0.5 M NH4Cl in stimulating ribosomes for AcPhe-tRNA and fMet-tRNA binding, for the puromycin reaction, and for incorporating acetylphenylalanine from AcPhe-tRNA into polypeptide. The factors appear to differ from the initiation factors, the elongation factor EF-T, and ribosomal proteins. Some uncertainty exists as to whether factor B is different from EF-G. To maximize the effect of the factors in initiator tRNA binding, we preincubated the ribosomes with the factors and carried out the binding assay for a short period at 15 degrees C. Maximal stimulation of binding occurred after about a 2-min preincubation at 37 degrees C. Longer preincubation times were required at 15 degrees C, and only slight stimulation was observed after preincubation at 0 degrees C. The extent of stimulation by the factors was not affected when the NH4Cl concentration was increased from 40 to 500 mM in the preincubation. The presence of both the 30S and 50S ribosomal subunits is required for the enhancement of AcPhe-tRNA binding. Polyphenylalanine synthesis carried out without AcPhe-tRNA is inhibited by the factors. It is suggested that the factors may act by inducing a structural rearrangement of the ribosomes.
mesh_terms: D001426:Bacterial Proteins / Q000302:isolation & purification / Q000494:pharmacology*; D004926:Escherichia coli / Q000378:metabolism*; D007700:Kinetics; D008715:Methionine / Q000378:metabolism; D010452:Peptide Biosynthesis; D010442:Peptide Chain Initiation, Translational / Q000187:drug effects*; D010445:Peptide Elongation Factors / Q000302:isolation & purification; D010448:Peptide Initiation Factors / Q000302:isolation & purification; D010649:Phenylalanine / Q000031:analogs & derivatives / Q000378:metabolism; D011691:Puromycin / Q000378:metabolism; D012343:RNA, Transfer / Q000378:metabolism; D012270:Ribosomes / Q000378:metabolism
chemical_list: D001426:Bacterial Proteins; D010445:Peptide Elongation Factors; D010448:Peptide Initiation Factors; D010649:Phenylalanine; D011691:Puromycin; D012343:RNA, Transfer; D008715:Methionine