The conserved Lysine69 residue plays a catalytic role in Mycobacterium tuberculosis shikimate dehydrogenase.

BMC Research Notes
Valnês da Silva RodriguesLuiz Augusto Basso

Abstract

The shikimate pathway is an attractive target for the development of antitubercular agents because it is essential in Mycobacterium tuberculosis, the causative agent of tuberculosis, but absent in humans. M. tuberculosis aroE-encoded shikimate dehydrogenase catalyzes the forth reaction in the shikimate pathway. Structural and functional studies indicate that Lysine69 may be involved in catalysis and/or substrate binding in M. tuberculosis shikimate dehydrogenase. Investigation of the kinetic properties of mutant enzymes can bring important insights about the role of amino acid residues for M. tuberculosis shikimate dehydrogenase. We have performed site-directed mutagenesis, steady-state kinetics, equilibrium binding measurements and molecular modeling for both the wild-type M. tuberculosis shikimate dehydrogenase and the K69A mutant enzymes. The apparent steady-state kinetic parameters for the M. tuberculosis shikimate dehydrogenase were determined; the catalytic constant value for the wild-type enzyme (50 s-1) is 68-fold larger than that for the mutant K69A (0.73 s-1). There was a modest increase in the Michaelis-Menten constant for DHS (K69A = 76 microM; wild-type = 29 microM) and NADPH (K69A = 30 microM; wild-type = 11 micro...Continue Reading

References

Jan 1, 1990·Critical Reviews in Biochemistry and Molecular Biology·R Bentley
Feb 1, 1995·Protein Engineering·A C WallaceJ M Thornton
Sep 1, 1994·Protein Science : a Publication of the Protein Society·A Sali, J P Overington
Dec 5, 1993·Journal of Molecular Biology·A Sali, T L Blundell
Nov 5, 1999·Advances in Experimental Medicine and Biology·L A Basso, J S Blanchard
Aug 15, 2000·Annual Review of Biophysics and Biomolecular Structure·M A Martí-RenomA Sali
Oct 3, 2002·Protein Expression and Purification·Maria L B MagalhãesDiógenes S Santos
May 8, 2004·Trends in Plant Science·Brandon d Moore
Dec 15, 2004·The Journal of Biological Chemistry·Holger A LindnerMiroslaw Cygler
Mar 1, 2005·The Journal of Biological Chemistry·Sasha SinghDinesh Christendat
Apr 13, 2005·Biochemistry·James G Robertson
Oct 8, 2005·Journal of Computational Chemistry·David Van Der SpoelHerman J C Berendsen
Nov 22, 2005·Protein Expression and Purification·Isabel O FonsecaLuiz A Basso
Nov 1, 2006·Protein Science : a Publication of the Protein Society·Min-Yi Shen, Andrej Sali
Dec 21, 2006·Archives of Biochemistry and Biophysics·Isabel O FonsecaDiógenes S Santos
Mar 14, 2007·Current Drug Targets·R G DucatiD S Santos
Sep 22, 2007·Current Opinion in Structural Biology·James G Robertson
Feb 9, 2008·Proteins·Helen A ArcuriWalter F de Azevedo
May 26, 2009·International Journal of Biological Macromolecules·Valnês da Silva RodriguesDiógenes Santiago Santos

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Methods Mentioned

BETA
PCR
X-ray

Software Mentioned

VERIFY 3D
MODELLER9v1
PROCHECK
SigmaPlot
LIGPLOT
GROMACS
MODELLER

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