1F8W

CRYSTAL STRUCTURE OF NADH PEROXIDASE MUTANT: R303M

Summary for 1F8W

• Entry DOI: 10.2210/pdb1f8w/pdb
• Related: 1JOA 1NHP 1NHQ 1NHR 1NHS 1NPX
• Descriptor: NADH PEROXIDASE, FLAVIN-ADENINE DINUCLEOTIDE (3 entities in total)
• Functional Keywords: interface, fad, nad-binding domains, oxidoreductase
• Biological source: Enterococcus faecalis
• Total number of polymer chains: 1
• Total formula weight: 50378.78

Authors

Yeh, J.I.,Claiborne, A.,Hol, W.G.J. (deposition date: 2000-07-05, release date: 2001-02-05, Last modification date: 2021-11-03)

Primary citation

Crane III, E.J.,Yeh, J.I.,Luba, J.,Claiborne, A.
Analysis of the kinetic and redox properties of the NADH peroxidase R303M mutant: correlation with the crystal structure.
Biochemistry, 39:10353-10364, 2000

PubMed 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.

PubMed: 10956025
DOI: 10.1021/bi000553m

Experimental method

X-RAY DIFFRACTION (2.45 Å)