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Analysis of the kinetic and redox properties of the NADH peroxidase R303M mutant: correlation with the crystal structure.

Crane EJ, Yeh JI, Luba J, Claiborne A
Biochemistry 39 10353-64 (2000)
Cited: 4 times
EuropePMC logo PMID: 10956025

Abstract

The crystal structure of the flavoprotein NADH peroxidase shows that the Arg303 side chain forms a hydrogen bond with the active-site His10 imidazole and is therefore likely to influence the catalytic mechanism. Dithionite titration of an R303M mutant [E(FAD, Cys42-sulfenic acid)] yields a two-electron reduced intermediate (EH(2)) with enhanced flavin fluorescence and almost no charge-transfer absorbance at pH 7.0; the pK(a) for the nascent Cys42-SH is increased by over 3.5 units in comparison with the wild-type EH(2) pK(a) of Cys42-SOH. The crystal structure of the R303M peroxidase has been refined at 2.45 A resolution. In addition to eliminating the Arg303 interactions with His10 and Glu14, the mutant exhibits a significant change in the conformation of the Cys42-SOH side chain relative to FAD and His10 in particular. These and other results provide a detailed understanding of Arg303 and its role in the structure and mechanism of this unique flavoprotein peroxidase.

Articles - 1f8w mentioned but not cited (1)

  1. Crystallization and preliminary analysis of a water-forming NADH oxidase from Lactobacillus sanfranciscensis. Lountos GT, Riebel BR, Wellborn WB, Bommarius AS, Orville AM. Acta Crystallogr D Biol Crystallogr 60 2044-2047 (2004)


Reviews citing this publication (1)

  1. The physiological role of reversible methionine oxidation. Drazic A, Winter J. Biochim Biophys Acta 1844 1367-1382 (2014)

Articles citing this publication (2)

  1. A hemin/G-quadruplex acts as an NADH oxidase and NADH peroxidase mimicking DNAzyme. Golub E, Freeman R, Willner I. Angew Chem Int Ed Engl 50 11710-11714 (2011)
  2. E. coli methionine sulfoxide reductase with a truncated N terminus or C terminus, or both, retains the ability to reduce methionine sulfoxide. Boschi-Muller S, Azza S, Branlant G. Protein Sci 10 2272-2279 (2001)


Related citations provided by authors (1)

  1. Structure of the native cysteine-sulfenic acid redox center of enterococcal NADH peroxidase refined at 2.8 A resolution.. Yeh JI, Claiborne A, Hol WG Biochemistry 35 9951-7 (1996)

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