Biochemical and physiological properties of the DNA binding domain of AraC protein

Journal of Molecular Biology
Andrew TimmesRobert Schleif

Abstract

Intact AraC protein is poorly soluble and difficult to purify, whereas its dimerization domain is the opposite. Unexpectedly, the DNA binding domain of AraC proved also to be soluble in cells when overproduced and is easily purified to homogeneity. The DNA binding affinity of the DNA binding domain for its binding site could not be measured by electrophoretic mobility shift because of its rapid association and dissociation rates, but its affinity could be measured with a fluorescence assay and was found to have a dissociation constant of 1 x 10(-8)M in 100 mM KCl. The binding of monomers of the DNA binding domain to adjacent half-sites occurs without substantial positive or negative cooperativity. A simple analysis relates the DNA binding affinities of monomers of DNA binding domain and normal dimeric AraC protein.

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Citations

Apr 7, 2010·Journal of Bacteriology·Patricia Domínguez-CuevasSilvia Marqués
Feb 17, 2009·Journal of Bacteriology·Stephanie DirlaRobert Schleif
Feb 26, 2008·Journal of Bacteriology·Patricia Domínguez-CuevasSilvia Marqués
Aug 16, 2011·Journal of Bacteriology·Jennifer Seedorff, Robert Schleif
Oct 5, 2011·Molecular and Cellular Biology·Jeffrey N McKnightGregory D Bowman
Jun 8, 2010·BMC Systems Biology·Carla J DavidsonMichael G Surette
Jul 12, 2008·PLoS Computational Biology·Namiko MitaraiKim Sneppen
Sep 10, 2013·Journal of Bacteriology·Ji Zeng, Stephen Spiro
May 8, 2009·Proteins·Michael E Rodgers, Robert Schleif
Jun 19, 2008·Proteins·Michael E RodgersRobert Schleif
Mar 3, 2012·Proteins·Stephanie Dirla Cole, Robert Schleif
Jun 13, 2015·Journal of the Royal Society, Interface·Edward J HancockAntonis Papachristodoulou
May 13, 2008·Biophysical Journal·Judith A MegerleJoachim O Rädler
Apr 11, 2021·FEMS Microbiology Reviews·Daniel Cortés-AvalosJ Antonio Ibarra
Apr 28, 2021·Nature Chemical Biology·Edoardo RomanoBarbara Di Ventura
Nov 1, 2005·EcoSal Plus·Christoph Mayer, Winfried Boos
Jun 15, 2019·Biochemistry·Matthew J Brown, Robert Schleif
Aug 22, 2013·Biochemistry·Robert F Schleif

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