TY - JOUR
T1 - Tungsten-containing formate dehydrogenase from Desulfovibrio gigas: metal identification and preliminary structural data by multi-wavelength crystallography
AU - Raaijmakers, Hans
AU - Teixeira, Susana
AU - Dias, João Miguel
AU - Almendra, Maria João
AU - Brondino, Carlos D.
AU - Moura, Isabel
AU - Moura, José J. G.
AU - Romão, Maria João
PY - 2001/1/1
Y1 - 2001/1/1
N2 - 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.
AB - 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.
KW - Formate dehydrogenase
KW - Iron-sulfur centers
KW - Molybdopterin
KW - Multi-wavelength anomalous dispersion
KW - Tungsten-containing enzymes
UR - http://www.scopus.com/inward/record.url?scp=0034852815&partnerID=8YFLogxK
U2 - 10.1007/s007750100215
DO - 10.1007/s007750100215
M3 - Article
C2 - 11372198
AN - SCOPUS:0034852815
SN - 0949-8257
VL - 6
SP - 398
EP - 404
JO - JBIC Journal of Biological Inorganic Chemistry
JF - JBIC Journal of Biological Inorganic Chemistry
IS - 4
ER -