PMID:10471730
| Citation |
Ibba, M, Sever, S, Praetorius-Ibba, M and Söll, D (1999) Transfer RNA identity contributes to transition state stabilization during aminoacyl-tRNA synthesis. Nucleic Acids Res. 27:3631-7 |
|---|---|
| Abstract |
Sequence-specific interactions between aminoacyl-tRNA synthetases and their cognate tRNAs ensure both accurate RNA recognition and the efficient catalysis of aminoacylation. The effects of tRNA(Trp)variants on the aminoacylation reaction catalyzed by wild-type Escherichia coli tryptophanyl-tRNA synthe-tase (TrpRS) have now been investigated by stopped-flow fluorimetry, which allowed a pre-steady-state analysis to be undertaken. This showed that tRNA(Trp)identity has some effect on the ability of tRNA to bind the reaction intermediate TrpRS-tryptophanyl-adenylate, but predominantly affects the rate at which trypto-phan is transferred from TrpRS-tryptophanyl adenylate to tRNA. Use of the binding ( K (tRNA)) and rate constants ( k (4)) to determine the energetic levels of the various species in the aminoacylation reaction showed a difference of approximately 2 kcal mol(-1)in the barrier to transition state formation compared to wild-type for both tRNA(Trp)A-->C73 and. These results directly show that tRNA identity contributes to the degree of complementarity to the transition state for tRNA charging in the active site of an aminoacyl-tRNA synthetase:aminoacyl-adenylate:tRNA complex. |
| Links | |
| Keywords |
Adenosine Monophosphate/metabolism; Base Sequence; Binding Sites; Catalysis; Escherichia coli/enzymology; Escherichia coli/genetics; Fluorescence; Kinetics; Mutation; Nucleic Acid Conformation; RNA, Transfer, Amino Acyl/biosynthesis; RNA, Transfer, Amino Acyl/chemistry; RNA, Transfer, Amino Acyl/metabolism; RNA, Transfer, Gln/chemistry; RNA, Transfer, Gln/genetics; RNA, Transfer, Gln/metabolism; RNA, Transfer, Trp/chemistry; RNA, Transfer, Trp/genetics; RNA, Transfer, Trp/metabolism; Substrate Specificity; Thermodynamics; Tryptophan/metabolism; Tryptophan-tRNA Ligase/metabolism |
| edit table |
Significance
You can help EcoliWiki by summarizing why this paper is useful
Useful Materials and Methods
You can help Ecoliwiki by describing the useful materials (strains, plasmids, antibodies, etc) described in this paper.
Annotations
<annotationlinks/>
EcoliWiki Links
Add links to pages that link here (e.g. gene, product, method pages)
References
See Help:References for how to manage references in EcoliWiki.