Document Type
Article
Publication Date
9-3-2013
Abstract
We have cloned genes encoding elongation factors We have cloned genes encoding elongation factors EF-Tu and EF-Ts from Pseudomonas aeruginosa and expressed and purified the proteins to greater than 95% homogeneity. Sequence analysis indicated that P. aeruginosa EF-Tu and EF-Ts are 84% and 55% identical to E. coli counterparts, respectively. P. aeruginosa EF-Tu was active when assayed in GDP exchange assays. Kinetic parameters for the interaction of EF-Tu with GDP in the absence of EF-Ts were observed to be = 33 μM, = 0.003 s−1, and the specificity constant was s−1 μM−1. In the presence of EF-Ts, these values were shifted to = 2 μM, = 0.005 s−1, and the specificity constant was s−1 μM−1. The equilibrium dissociation constants governing the binding of EF-Tu to GDP () were 30–75 nM and to GTP () were 125–200 nM. EF-Ts stimulated the exchange of GDP by EF-Tu 10-fold. P. aeruginosa EF-Tu was active in forming a ternary complex with GTP and aminoacylated tRNA and was functional in poly(U)-dependent binding of Phe-tRNAPhe at the A-site of P. aeruginosa ribosomes. P. aeruginosa EF-Tu was active in poly(U)-programmed polyphenylalanine protein synthesis system composed of all P. aeruginosa components.
Recommended Citation
Palmer, Stephanie O., Edna Y. Rangel, Alberto E. Montalvo, Alexis T. Tran, Kate C. Ferguson, and James M. Bullard. "Cloning and Characterization of EF‐Tu and EF‐Ts from Pseudomonas aeruginosa." BioMed research international 2013, no. 1 (2013): 585748. https://doi.org/10.1155/2013/585748
Creative Commons License
This work is licensed under a Creative Commons Attribution 3.0 License.
Publication Title
BioMed Research International
DOI
10.1155/2013/585748
Comments
© 2013 Hindawi Publishing Corporation. Published under Creative Commons Attribution License. Original published version available at http://doi.org/10.1155/2013/585748.