Tungsten-containing formate dehydrogenase from Desulfovibrio gigas: metal identification and preliminary structural data by multi-wavelength crystallography

Hans Raaijmakers, Susana Teixeira, João Miguel Dias, Maria João Almendra, Carlos D. Brondino, Isabel Moura, José J. G. Moura, Maria João Romão

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38 Citations (Scopus)

Abstract

The tungsten-containing formate dehydrogenase (W-FDH) isolated from Desulfovibrio gigas has been crystallized in space group P21, with cell parameters a=73.8 Å, b=111.3 Å, c=156.6 Å and β=93.7°. These crystals diffract to beyond 2.0 Å on a synchrotron radiation source. W-FDH is a heterodimer (92 kDa and 29 kDa subunits) and two W-FDH molecules are present in the asymmetric unit. Although a molecular replacement solution was found using the periplasmic nitrate reductase as a search model, additional phasing information was needed. A multiple-wavelength anomalous dispersion (MAD) dataset was collected at the W- and Fe-edges, at four different wavelengths. Anomalous and dispersive difference data allowed us to unambiguously identify the metal atoms bound to W-FDH as one W atom with a Se-cysteine ligand as well as one [4Fe-4S] cluster in the 92 kDa subunit, and three additional [4Fe-4S] centers in the smaller 29 kDa subunit. The D. gigas W-FDH was previously characterized based on metal analysis and spectroscopic data. One W atom was predicted to be bound to two molybdopterin guanine dinucleotide (MGD) pterin cofactors and two [4Fe-4S] centers were proposed to be present. The crystallographic data now reported reveal a selenium atom (as a Se-cysteine) coordinating to the W site, as well as two extra [4Fe-4S] clusters not anticipated before. The EPR data were re-evaluated in the light of these new results.

Original languageEnglish
Pages (from-to)398-404
Number of pages7
JournalJournal of Biological Inorganic Chemistry
Volume6
Issue number4
DOIs
Publication statusPublished - 1 Jan 2001

Keywords

  • Formate dehydrogenase
  • Iron-sulfur centers
  • Molybdopterin
  • Multi-wavelength anomalous dispersion
  • Tungsten-containing enzymes

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