title: Contribution of the esterified amino acid to the binding of aminoacylated tRNAs to the ribosomal P- and A-sites.
abstract: Crystallographic studies suggest that the esterified amino acid of aminoacyl tRNA make contacts with the ribosomal A-site but not in the P-site. Biochemical evidence indicating a thermodynamic contribution of the esterified amino acid to binding aminoacyl-tRNA to either the ribosomal P- and A-sites has been inconsistent, partly because of the labile nature of the aminoacyl linkage and the long times required to reach equilibrium. Measuring the association and dissociation rates of deacylated and aminoacylated tRNAs to the A-site and P-site of E. coli ribosomes afforded an accurate estimate of the contribution of the amino acid. While esterified phenylalanine or methionine has no effect on the affinity of tRNA to the P-site, an esterified pheylalanine stabilizes binding to the A-site by 7 kJ/mol, in agreement with the contacts observed in the X-ray crystal structure. In addition, it was shown that the presence of an esterified amino acid in one ribosomal site does not affect the binding of an aa-tRNA to the other site.
authors.affiliation: The Department of Biochemistry, Molecular Biology & Cell Biology, Northwestern University, 2205 Tech Dr., 2-100 Hogan Hall, Evanston, Illinois 60208, USA.
authors.forename: Richard P
authors.initials: RP
authors.lastname: Fahlman
authors.forename: Olke C
authors.initials: OC
authors.lastname: Uhlenbeck
mesh_terms: D000215:Acylation; D000596:Amino Acids / Q000737:chemistry* / Q000378:metabolism*; D001483:Base Sequence; D001665:Binding Sites; D004951:Esterification; D007700:Kinetics; D008958:Models, Molecular; D009690:Nucleic Acid Conformation; D012346:RNA, Transfer, Amino Acyl / Q000737:chemistry / Q000378:metabolism*; D012358:RNA, Transfer, Met / Q000737:chemistry / Q000378:metabolism; D012360:RNA, Transfer, Phe / Q000378:metabolism; D012270:Ribosomes / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D000596:Amino Acids; D012346:RNA, Transfer, Amino Acyl; D012358:RNA, Transfer, Met; D012360:RNA, Transfer, Phe