Purification and preliminary characterization of tetraheme cytochrome c3 and adenylylsulfate reductase from the peptidolytic sulfate-reducing bacterium Desulfovibrio aminophilus DSM 12254

Alejandro López-Cortés, Sergey Bursakov, Angelo Figueiredo, Anders E. Thapper, Smilja Todorovic, José J. G. Moura, Bernard Ollivier, Isabel Moura, Guy Fauque

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Abstract

Two proteins were purified and preliminarily characterized from the soluble extract of cells (310 g, wet weight) of the aminolytic and peptidolytic sulfate-reducing bacterium Desulfovibrio (D.) aminophilus DSM12254. The iron-sulfur flavoenzyme adenylylsulfate (adenosine 5′-phosphosulfate, APS) reductase, a key enzyme in the microbial dissimilatory sulfate reduction, has been purified in three chromatographic steps(DEAE-Biogel A, Source 15, and Superdex 200 columns). It contains two different subunits with molecular masses of 75 and 18 kDa. The fraction after the last purification step had a purity index ( A 278 nm / A 388 nm) of5.34, which was used for further EPR spectroscopic studies. The D. aminophilus APS reductase is very similar to the homologous enzymes isolated from D. gigas and D. desulfuricans ATCC 27774. A tetrahemecytochrome c3 (His-heme iron-His) has been purified in three chromatographic steps (DEAE- Biogel A,Source 15, and Biogel-HTP columns) and preliminarily characterized. It has a purity index ( [ A 553 nm - A 570 nm] red / A 280nm ox) of 2.9 and a molecular mass of around 15 kDa, and its spectroscopic characterization(NMR and EPR) has been carried out. This hemoprotein presents similarities with the tetraheme cytochrome c3 from Desulfomicrobium (Des.) norvegicum (NMR spectra, and N-terminal amino acid sequence).

Original languageEnglish
Pages (from-to)81-91
Number of pages11
JournalBioinorganic Chemistry And Applications
Volume2005
Issue number1-2
DOIs
Publication statusPublished - 1 Dec 2005

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Desulfovibrio
Cytochrome Reductases
Molecular mass
4 alpha-glucanotransferase
Sulfates
Purification
Paramagnetic resonance
Bacteria
Iron
Nuclear magnetic resonance
Enzymes
Cell Extracts
Heme
Sulfur
Amino Acid Sequence
Amino Acids
Weights and Measures
Proteins
adenylylsulfate reductase
cytochrome c(3)

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@article{181d02190e034bff9fd5b8436b8997ad,
title = "Purification and preliminary characterization of tetraheme cytochrome c3 and adenylylsulfate reductase from the peptidolytic sulfate-reducing bacterium Desulfovibrio aminophilus DSM 12254",
abstract = "Two proteins were purified and preliminarily characterized from the soluble extract of cells (310 g, wet weight) of the aminolytic and peptidolytic sulfate-reducing bacterium Desulfovibrio (D.) aminophilus DSM12254. The iron-sulfur flavoenzyme adenylylsulfate (adenosine 5′-phosphosulfate, APS) reductase, a key enzyme in the microbial dissimilatory sulfate reduction, has been purified in three chromatographic steps(DEAE-Biogel A, Source 15, and Superdex 200 columns). It contains two different subunits with molecular masses of 75 and 18 kDa. The fraction after the last purification step had a purity index ( A 278 nm / A 388 nm) of5.34, which was used for further EPR spectroscopic studies. The D. aminophilus APS reductase is very similar to the homologous enzymes isolated from D. gigas and D. desulfuricans ATCC 27774. A tetrahemecytochrome c3 (His-heme iron-His) has been purified in three chromatographic steps (DEAE- Biogel A,Source 15, and Biogel-HTP columns) and preliminarily characterized. It has a purity index ( [ A 553 nm - A 570 nm] red / A 280nm ox) of 2.9 and a molecular mass of around 15 kDa, and its spectroscopic characterization(NMR and EPR) has been carried out. This hemoprotein presents similarities with the tetraheme cytochrome c3 from Desulfomicrobium (Des.) norvegicum (NMR spectra, and N-terminal amino acid sequence).",
author = "Alejandro L{\'o}pez-Cort{\'e}s and Sergey Bursakov and Angelo Figueiredo and Thapper, {Anders E.} and Smilja Todorovic and Moura, {Jos{\'e} J. G.} and Bernard Ollivier and Isabel Moura and Guy Fauque",
year = "2005",
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T1 - Purification and preliminary characterization of tetraheme cytochrome c3 and adenylylsulfate reductase from the peptidolytic sulfate-reducing bacterium Desulfovibrio aminophilus DSM 12254

AU - López-Cortés, Alejandro

AU - Bursakov, Sergey

AU - Figueiredo, Angelo

AU - Thapper, Anders E.

AU - Todorovic, Smilja

AU - Moura, José J. G.

AU - Ollivier, Bernard

AU - Moura, Isabel

AU - Fauque, Guy

PY - 2005/12/1

Y1 - 2005/12/1

N2 - Two proteins were purified and preliminarily characterized from the soluble extract of cells (310 g, wet weight) of the aminolytic and peptidolytic sulfate-reducing bacterium Desulfovibrio (D.) aminophilus DSM12254. The iron-sulfur flavoenzyme adenylylsulfate (adenosine 5′-phosphosulfate, APS) reductase, a key enzyme in the microbial dissimilatory sulfate reduction, has been purified in three chromatographic steps(DEAE-Biogel A, Source 15, and Superdex 200 columns). It contains two different subunits with molecular masses of 75 and 18 kDa. The fraction after the last purification step had a purity index ( A 278 nm / A 388 nm) of5.34, which was used for further EPR spectroscopic studies. The D. aminophilus APS reductase is very similar to the homologous enzymes isolated from D. gigas and D. desulfuricans ATCC 27774. A tetrahemecytochrome c3 (His-heme iron-His) has been purified in three chromatographic steps (DEAE- Biogel A,Source 15, and Biogel-HTP columns) and preliminarily characterized. It has a purity index ( [ A 553 nm - A 570 nm] red / A 280nm ox) of 2.9 and a molecular mass of around 15 kDa, and its spectroscopic characterization(NMR and EPR) has been carried out. This hemoprotein presents similarities with the tetraheme cytochrome c3 from Desulfomicrobium (Des.) norvegicum (NMR spectra, and N-terminal amino acid sequence).

AB - Two proteins were purified and preliminarily characterized from the soluble extract of cells (310 g, wet weight) of the aminolytic and peptidolytic sulfate-reducing bacterium Desulfovibrio (D.) aminophilus DSM12254. The iron-sulfur flavoenzyme adenylylsulfate (adenosine 5′-phosphosulfate, APS) reductase, a key enzyme in the microbial dissimilatory sulfate reduction, has been purified in three chromatographic steps(DEAE-Biogel A, Source 15, and Superdex 200 columns). It contains two different subunits with molecular masses of 75 and 18 kDa. The fraction after the last purification step had a purity index ( A 278 nm / A 388 nm) of5.34, which was used for further EPR spectroscopic studies. The D. aminophilus APS reductase is very similar to the homologous enzymes isolated from D. gigas and D. desulfuricans ATCC 27774. A tetrahemecytochrome c3 (His-heme iron-His) has been purified in three chromatographic steps (DEAE- Biogel A,Source 15, and Biogel-HTP columns) and preliminarily characterized. It has a purity index ( [ A 553 nm - A 570 nm] red / A 280nm ox) of 2.9 and a molecular mass of around 15 kDa, and its spectroscopic characterization(NMR and EPR) has been carried out. This hemoprotein presents similarities with the tetraheme cytochrome c3 from Desulfomicrobium (Des.) norvegicum (NMR spectra, and N-terminal amino acid sequence).

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U2 - 10.1155/BCA.2005.81

DO - 10.1155/BCA.2005.81

M3 - Article

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EP - 91

JO - Bioinorganic Chemistry And Applications

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SN - 1565-3633

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