Substrate-assisted cysteine deprotonation in the mechanism of dimethylargininase (DDAH) from Pseudomonas aeruginosa

Biochemistry
Everett M StoneWalter Fast

Abstract

The enzyme dimethylargininase (also known as dimethylarginine dimethylaminohydrolase or DDAH; EC 3.5.3.18) catalyzes the hydrolysis of endogenous nitric oxide synthase inhibitors, N(omega)-methyl-l-arginine and N(omega),N(omega)-dimethyl-l-arginine. Understanding the mechanism and regulation of DDAH activity is important for developing ways to control nitric oxide production during angiogenesis and in many cases of vascular endothelial pathobiology. Several possible physiological regulation mechanisms of DDAH depend upon the presence of an active-site cysteine residue, Cys249 in Pseudomonas aeruginosa (Pa) DDAH, which is proposed to serve as a nucleophile in the catalytic mechanism. Through the use of pH-dependent ultraviolet and visible (UV-vis) difference spectroscopy and inactivation kinetics, the pK(a) of the active-site Cys249 in the resting enzyme was found to be unperturbed from pK(a) values of typical noncatalytic cysteine residues. In contrast, the pH dependence of k(cat) values indicates a much lower apparent pK(a) value. UV-vis difference spectroscopy between wild-type and C249S DDAH shows absorbance changes consistent with Cys249 deprotonation to the anionic thiolate upon binding positively charged ligands. The prot...Continue Reading

References

Jan 1, 1993·Methods in Enzymology·L M UtschigT V O'Halloran
Jul 2, 2002·The Journal of Biological Chemistry·Arturo J Cardounel, Jay L Zweier
Sep 19, 2002·British Journal of Cancer·V KostourouG St J Whitley
Nov 21, 2002·The Journal of Biological Chemistry·Markus KnippMilan Vasák
Dec 4, 2002·Nature Reviews. Drug Discovery·Patrick Vallance, James Leiper
Apr 27, 2004·The Journal of Biological Chemistry·Anssi M MalinenReijo Lahti
Feb 24, 2005·Journal of the American Chemical Society·Markus KnippMilan Vasák
Jun 1, 2005·Vascular Medicine·V AchanJ P Cooke

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Citations

Oct 14, 2010·Biochemistry·Justin E JonesPaul R Thompson
May 15, 2007·Biochemistry·Bryan KnuckleyPaul R Thompson
Jan 13, 2011·Journal of the American Chemical Society·Corey M JohnsonWalter Fast
Jun 3, 2011·Journal of the American Chemical Society·Corey M JohnsonWalter Fast
Aug 12, 2009·Journal of the American Chemical Society·Grigory SmolentsevPieter Glatzel
Sep 30, 2008·Drug Metabolism Reviews·Young-Mi AhSang Geon Kim
May 15, 2015·Chemical Reviews·Jakob FuhrmannPaul R Thompson
Sep 21, 2010·Archives of Biochemistry and Biophysics·Sofía B RodríguezDavid E Ash
Jul 27, 2010·Biochimica Et Biophysica Acta·Thomas Linsky, Walter Fast
Dec 29, 2009·Bioorganic Chemistry·Justin E JonesPaul R Thompson
Jun 30, 2007·Protein Science : a Publication of the Protein Society·Oliver BraunMilan Vasák
Dec 17, 2009·Chembiochem : a European Journal of Chemical Biology·Bryan KnuckleyPaul R Thompson
Nov 27, 2014·Journal of Molecular Graphics & Modelling·Dongmei LiJianping Lin
May 17, 2008·Chemistry & Biology·Thomas W LinskyWalter Fast
May 18, 2016·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Rhys B MurphyArduino A Mangoni
Sep 21, 2007·American Journal of Physiology. Cell Physiology·Arthur J PopeArturo J Cardounel
Apr 2, 2016·The Biochemical Journal·Charles A Soares, Bryan Knuckley
Oct 16, 2007·American Journal of Physiology. Heart and Circulatory Physiology·Fredrik PalmChristopher S Wilcox

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