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.

Comments

© 2013 Hindawi Publishing Corporation. Published under Creative Commons Attribution License. Original published version available at http://doi.org/10.1155/2013/585748.

Palmer, S., Rangel, E., Montalvo, R., et al. Cloning and Characterization of EF-Tu and EF-Ts From Pseudomonas Aeruginosa. BioMed Research International 2013, vol. 2013, article no. 585748.

Publication Title

BioMed Research International

DOI

10.1155/2013/585748

Included in

Chemistry Commons

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