Purification and characterization of a tungsten-containing formate dehydrogenase from Desulfovibrio gigas

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Abstract

An air-stable formate dehydrogenase (FDH), an enzyme that catalyzes the oxidation of formate to carbon dioxide, was purified from the sulfate reducing organism Desulfovibrio gigas (D. gigas) NCIB 9332. D. gigas FDH is a heterodimeric protein [α (92 kDa) and β (29 kDa) subunits] and contains. 7 ± 1 Fe/protein and 0.9 ± 0.1 W/protein. Selenium was not detected. The UV/visible absorption spectrum of D. gigas FDH is typical of an iron-sulfur protein. Analysis of pterin nucleotides yielded a content of 1.3 ± 0.1 guanine monophosphate/mol of enzyme, which suggests a tungsten coordination with two molybdopterin guanine dinucleotide cofactors. Both Mossbauer spectroscopy performed on D. gigas FDH grown in a medium enriched with 57Fe and EPR studies performed in the native and fully reduced state of the protein confirmed the presence of two [4Fe-4S] clusters. Variable-temperature EPR studies showed the presence of two signals compatible with an atom in a d1 configuration albeit with an unusual relaxation behavior as compared to the one generally observed for W(V) ions.

Original languageEnglish
Pages (from-to)16366-16372
Number of pages7
JournalBiochemistry
Volume38
Issue number49
DOIs
Publication statusPublished - 7 Dec 1999

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Desulfovibrio gigas
Formate Dehydrogenases
Tungsten
Purification
formic acid
Paramagnetic resonance
Proteins
Mossbauer Spectroscopy
Pterins
Iron-Sulfur Proteins
Guanine
Mossbauer spectroscopy
Enzymes
Selenium
Carbon Dioxide
Sulfates
Absorption spectra
Nucleotides
Air
Ions

Cite this

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title = "Purification and characterization of a tungsten-containing formate dehydrogenase from Desulfovibrio gigas",
abstract = "An air-stable formate dehydrogenase (FDH), an enzyme that catalyzes the oxidation of formate to carbon dioxide, was purified from the sulfate reducing organism Desulfovibrio gigas (D. gigas) NCIB 9332. D. gigas FDH is a heterodimeric protein [α (92 kDa) and β (29 kDa) subunits] and contains. 7 ± 1 Fe/protein and 0.9 ± 0.1 W/protein. Selenium was not detected. The UV/visible absorption spectrum of D. gigas FDH is typical of an iron-sulfur protein. Analysis of pterin nucleotides yielded a content of 1.3 ± 0.1 guanine monophosphate/mol of enzyme, which suggests a tungsten coordination with two molybdopterin guanine dinucleotide cofactors. Both Mossbauer spectroscopy performed on D. gigas FDH grown in a medium enriched with 57Fe and EPR studies performed in the native and fully reduced state of the protein confirmed the presence of two [4Fe-4S] clusters. Variable-temperature EPR studies showed the presence of two signals compatible with an atom in a d1 configuration albeit with an unusual relaxation behavior as compared to the one generally observed for W(V) ions.",
author = "Almendra, {Maria J.} and Brondino, {Carlos D.} and Olga Gavel and Pereira, {Alice S.} and Pedro Tavares and Sergey Bursakov and Rui Duarte and Jorge Caldeira and Moura, {Jos{\'e} J. G.} and Isabel Moura",
year = "1999",
month = "12",
day = "7",
doi = "10.1021/bi990069n",
language = "English",
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pages = "16366--16372",
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issn = "0006-2960",
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T1 - Purification and characterization of a tungsten-containing formate dehydrogenase from Desulfovibrio gigas

AU - Almendra, Maria J.

AU - Brondino, Carlos D.

AU - Gavel, Olga

AU - Pereira, Alice S.

AU - Tavares, Pedro

AU - Bursakov, Sergey

AU - Duarte, Rui

AU - Caldeira, Jorge

AU - Moura, José J. G.

AU - Moura, Isabel

PY - 1999/12/7

Y1 - 1999/12/7

N2 - An air-stable formate dehydrogenase (FDH), an enzyme that catalyzes the oxidation of formate to carbon dioxide, was purified from the sulfate reducing organism Desulfovibrio gigas (D. gigas) NCIB 9332. D. gigas FDH is a heterodimeric protein [α (92 kDa) and β (29 kDa) subunits] and contains. 7 ± 1 Fe/protein and 0.9 ± 0.1 W/protein. Selenium was not detected. The UV/visible absorption spectrum of D. gigas FDH is typical of an iron-sulfur protein. Analysis of pterin nucleotides yielded a content of 1.3 ± 0.1 guanine monophosphate/mol of enzyme, which suggests a tungsten coordination with two molybdopterin guanine dinucleotide cofactors. Both Mossbauer spectroscopy performed on D. gigas FDH grown in a medium enriched with 57Fe and EPR studies performed in the native and fully reduced state of the protein confirmed the presence of two [4Fe-4S] clusters. Variable-temperature EPR studies showed the presence of two signals compatible with an atom in a d1 configuration albeit with an unusual relaxation behavior as compared to the one generally observed for W(V) ions.

AB - An air-stable formate dehydrogenase (FDH), an enzyme that catalyzes the oxidation of formate to carbon dioxide, was purified from the sulfate reducing organism Desulfovibrio gigas (D. gigas) NCIB 9332. D. gigas FDH is a heterodimeric protein [α (92 kDa) and β (29 kDa) subunits] and contains. 7 ± 1 Fe/protein and 0.9 ± 0.1 W/protein. Selenium was not detected. The UV/visible absorption spectrum of D. gigas FDH is typical of an iron-sulfur protein. Analysis of pterin nucleotides yielded a content of 1.3 ± 0.1 guanine monophosphate/mol of enzyme, which suggests a tungsten coordination with two molybdopterin guanine dinucleotide cofactors. Both Mossbauer spectroscopy performed on D. gigas FDH grown in a medium enriched with 57Fe and EPR studies performed in the native and fully reduced state of the protein confirmed the presence of two [4Fe-4S] clusters. Variable-temperature EPR studies showed the presence of two signals compatible with an atom in a d1 configuration albeit with an unusual relaxation behavior as compared to the one generally observed for W(V) ions.

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