PMID:9662461
| Citation |
Mansilla, F, Knudsen, CR and Clark, BF (1998) Mutational analysis of Glu272 in elongation factor 1A of E. coli. FEBS Lett. 429:417-20 |
|---|---|
| Abstract |
In our previous work (Mansilla et al. (1997) Protein Eng. 10, 927-934) we showed that Arg7 of Escherichia coli elongation factor Tu (EF1A) plays an essential role in aminoacyl-tRNA (aa-tRNA) binding. Substitution of Arg7 by Ala or Glu lost this activity. We proposed that Arg7 forms a salt bridge with the charged conserved amino acid Glu272 (Asp284 in Thermus aquaticus) thereby binding the N-terminal region of the protein to domain 2 and thus completing the conformational rearrangement needed for binding aa-tRNA. In this work we have mutated Glu272 to arginine, either alone (Glu272Arg), or in combination with one of the above mentioned mutations (Arg7Glu/Glu272Arg) in order to test this hypothesis. Our results show that, in confirmation of our thesis based on structural knowledge, the substitution of Glu272 (Asp284) decreases the ability of EF1A:GTP to bind aa-tRNA. |
| Links | |
| Keywords |
Bacterial Proteins/genetics; Bacterial Proteins/metabolism; DNA Mutational Analysis; Escherichia coli; Glutamic Acid; Guanosine Diphosphate/metabolism; Guanosine Triphosphate/metabolism; Mutagenesis, Site-Directed; Peptide Chain Elongation, Translational; Peptide Elongation Factor Tu/genetics; Peptide Elongation Factor Tu/metabolism; RNA, Transfer, Amino Acyl/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.