Biophysical characterization of the TraY protein of Escherichia coli F factor.

The Journal of Biological Chemistry
J F SchildbachR T Sauer

Abstract

The TraY protein is required for efficient bacterial conjugation by Escherichia coli F factor. TraY has two functional roles: participating in the "relaxosome," a protein-DNA complex that nicks one strand of the F factor plasmid, and up-regulating transcription from the traYI promoter. The traY gene was cloned, and the TraY protein was expressed, purified, and characterized. TraY has a mixed alpha-helix and beta-sheet secondary structure as judged by its circular dichroism spectrum, is monomeric, and undergoes reversible urea denaturation with delta Gu = 6 kcal/mol at 25 degrees C. The kinetics of protein unfolding and refolding, as measured by changes in fluorescence, are complex, suggesting the presence of intermediates or of heterogeneity in the folding reaction. TraY has been classified as a member of the ribbon-helix-helix family of transcription factors but is unusual in appearing to have tandem repeats of the beta alpha alpha motif in the same polypeptide chain. The data presented here show that folding and assembly of the functional (beta alpha alpha)2 unit occurs as an intramolecular reaction and not by cross-folding between different polypeptide chains.

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Citations

Sep 11, 2002·Journal of Molecular Biology·Pamela L LumJoel F Schildbach
Mar 15, 2003·Biochimica Et Biophysica Acta·Lara M StreetJoel F Schildbach
Aug 7, 2007·Nature Reviews. Microbiology·Eric R Schreiter, Catherine L Drennan
Dec 14, 1999·Protein Science : a Publication of the Protein Society·P T Chivers, R T Sauer
Dec 31, 2008·Journal of Bacteriology·Athanasia VarsakiFernando de la Cruz
Jan 22, 2002·Journal of Molecular Biology·M M KrasilnikovaS M Mirkin
May 26, 2011·European Biophysics Journal : EBJ·Gundampati Ravi KumarMira Debnath
Jul 3, 1999·The Journal of Biological Chemistry·P L Lum, J F Schildbach
Nov 2, 1999·Biochemical and Biophysical Research Communications·S KunduM V Jagannadham
Sep 29, 2004·Biochimica Et Biophysica Acta·Gabriel Moncalián, Fernando de la Cruz

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