Biochemical, spectroscopic, and thermodynamic properties of flavodiiron proteins

Research output: Chapter in Book/Report/Conference proceedingChapter

42 Citations (Scopus)

Abstract

The flavodiiron proteins (FDPs), present in Archaea, Bacteria, and some protozoan pathogens (mostly anaerobes or microaerophiles), have been proposed to afford protection to microbes against nitric oxide and/or oxygen (toxic for anaerobes). The structural prototype of this protein family is a homodimer assembled in a "head‐to‐tail" configuration, with each monomer being composed of two domains: an N‐terminal metallo‐β‐lactamase module harboring a nonheme diiron center (active site of NO/O2 reduction) and a C‐terminal flavodoxin module, where a flavin mononucleotide moiety is embedded. Several FDPs bear C‐terminal extra domains, which influence the composition of the respective electron transfer chains that couple NAD(P)H oxidation to NO/O2 reduction. Herein are described methodologies employed to successfully produce, isolate, and characterize fully operative recombinant flavodiiron proteins. Spectroscopic techniques, namely absorption (visible and near‐ultraviolet) and electron paramagnetic resonance spectroscopies, allowed redox‐sensitive spectral fingerprints to be obtained, used further in the functional characterization of isolated flavodiiron proteins. Altogether, these studies on pure proteins contribute to understanding the molecular determinants that govern the in vivo function of the FDPs.
Original languageUnknown
Title of host publicationGlobins and Other Nitric Oxide-Reactive Proteins, Part B
EditorsRobert K. Poole
Place of PublicationSan Diego
PublisherElsevier Academic Press Inc
Pages21-45
Volume437
ISBN (Print)978-0-12-374278-0
DOIs
Publication statusPublished - 1 Jan 2008

Publication series

NameMethods in Enzymology
PublisherElsevier Academic Press Inc
ISSN (Print)0076-6879

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