PMID: 9182711Jun 1, 1997Paper

pH-dependence for binding a single nitrite ion to each type-2 copper centre in the copper-containing nitrite reductase of Alcaligenes xylosoxidans

The Biochemical Journal
Z H AbrahamR R Eady

Abstract

The first quantitative characterization of the interaction of NO2(-) with the Cu-containing dissimilatory nitrite reductase (NiR) of Alcaligenes xylosoxidans using steady-state kinetics, equilibrium gel filtration and EPR spectroscopy is described. Each molecule of this protein consists of three equivalent subunits, each containing a type-1 Cu atom and also a type-2 Cu atom at each subunit interface. Enzyme activity increased in a biphasic manner with decreasing pH, having an optimum at pH 5.2 and a plateau between pH 6.1 and 5.8. Equilibrium gel filtration showed that binding of NO2(-) to the oxidized NiR was also pH-dependent. At pH 7.5, no binding was detectable, but binding was detectable at lower pH values. At pH 5.2, the concentration-dependence for binding of NO2(-) to the enzyme showed that approx. 4.1 NO2(-) ions bound per trimeric NiR molecule. Unexpectedly, NiR deficient in type-2 Cu centres bound 1.3 NO2(-) ions per trimer. When corrected for this binding, a value of 3 NO2(-) ions bound per trimer of NiR, equivalent to the type-2 Cu content. The NO2(-)-induced changes in the EPR parameters of the type-2 Cu centre of the oxidized enzyme showed a similar pH-dependence to that of the activity. Binding constants for NO2...Continue Reading

Citations

Dec 7, 2006·The Journal of Biological Chemistry·Frida JacobsonRichard Neutze
Dec 3, 2013·Journal of Biochemistry·Yohta FukudaTsuyoshi Inoue
Jul 10, 2009·The Journal of Biological Chemistry·Sibylle BrennerNigel S Scrutton
Dec 20, 2017·World Journal of Microbiology & Biotechnology·Haofeng GaoNan Hu
Mar 16, 2018·Metallomics : Integrated Biometal Science·Masami Lintuluoto, Juha Mikael Lintuluoto
Apr 15, 2006·The Journal of Biological Chemistry·Hein J WijmaGerard W Canters
Jul 16, 2014·Nature Communications·Nicole G H LeferinkS Samar Hasnain
Feb 28, 2008·Proceedings of the National Academy of Sciences of the United States of America·Sofya KuznetsovaGerard W Canters
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May 20, 2016·Rapid Communications in Mass Spectrometry : RCM·M Alexandra WeigandDaniel M Sigman
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