The active centers of adenylylsulfate reductase from Desulfovibrio gigas Characterization and spectroscopic studies

Detalhes bibliográficos
Autor(a) principal: Lampreia, Jorge
Data de Publicação: 1990
Outros Autores: Moura, Isabel, Teixeira, Miguel, Peck, Harry D., LeGall, Jean, Huynh, Boi H., Moura, José J. G.
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: https://doi.org/10.1111/j.1432-1033.1990.tb15447.x
Resumo: NIGMS NIH HHS (GM 032187) NIDDK NIH HHS (DK 01135)
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spelling The active centers of adenylylsulfate reductase from Desulfovibrio gigas Characterization and spectroscopic studiesBiochemistryNIGMS NIH HHS (GM 032187) NIDDK NIH HHS (DK 01135)In order to utilize sulfate as the terminal electron acceptor, sulfate‐reducing bacteria are equipped with a complex enzymatic system in which adenylylsulfate (AdoPSO4) reductase plays one of the major roles, reducing AdoPSO4 (the activated form of sulfate) to sulfite, with release of AMP. The enzyme has been purified to homogeneity from the anaerobic sulfate reducer Desulfovibrio gigas. The protein is composed of two non‐identical subunits (70 kDa and 23 kDa) and is isolated in a multimeric form (∼ 400 kDa). It is an iron‐sulfur, flavincontaining protein, with one FAD moiety, eight iron atoms and a minimum molecular mass of 93 kDa. Low‐temperature EPR studies were performed to characterize its redox centers. In the native state, the enzyme showed an almost isotropic signal centered at g= 2.02 and only detectable below 20 K. This signal represented a minor species (0.10–0.25 spins/mol) and showed line broadening in the enzyme isolated from 57Fe‐grown cells. Addition of sulfite had a minor effect on the EPR spectrum, but caused a major decrease in the visible region of the optical spectrum (around 392 nm). Further addition of AMP induced only a minor change in the visible spectrum whereas major changes were seen in the EPR spectrum; the appearance of a rhombic signal at g values 2.096, 1.940 and 1.890 (reduced Fe‐S center I) observable below 30 K and a concomitant decrease in intensity of the g= 2.02 signal were detected. Effects of chemical reductants (ascorbate, H2/hydrogenase‐reduced methyl viologen and dithionite) were also studied. A short time reduction with dithionite (15 s) or reduction with methyl viologen gave rise to the full reduction of center I (with slightly modified g values at 2.079, 1.939 and 1.897), and the complete disappearance of the g= 2.02 signal. Further reduction with dithionite produces a very complex EPR spectrum of a spin–spin‐coupled nature (observable below 20 K), indicating the presence of at least two iron‐sulfur centers, (centers I and II). Mössbauer studies on 57Fe‐enriched D. gigas AdoPSO4 reductase demonstrated unambiguously the presence of two 4Fe clusters. Center II has a redox potential 400 mV and exhibits spectroscopic properties that are characteristic of a ferredoxin‐type [4Fe‐4S] cluster. Center I exhibits spectra with atypical Mössbauer parameters in its reduced state and has a midpoint potential around 0 mV, which is distinct from that of a ferredoxin‐type [4Fe‐4S] cluster, suggesting a different structure and/or a distinct cluster‐ligand environment.Instituto de Tecnologia Química e Biológica António Xavier (ITQB)RUNLampreia, JorgeMoura, IsabelTeixeira, MiguelPeck, Harry D.LeGall, JeanHuynh, Boi H.Moura, José J. G.2019-09-10T22:49:51Z1990-01-011990-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article12application/pdfhttps://doi.org/10.1111/j.1432-1033.1990.tb15447.xeng0014-2956PURE: 14640845http://www.scopus.com/inward/record.url?scp=0025320295&partnerID=8YFLogxKhttps://doi.org/10.1111/j.1432-1033.1990.tb15447.xinfo:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2024-03-11T04:35:45Zoai:run.unl.pt:10362/80775Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T03:35:56.067948Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse
dc.title.none.fl_str_mv The active centers of adenylylsulfate reductase from Desulfovibrio gigas Characterization and spectroscopic studies
title The active centers of adenylylsulfate reductase from Desulfovibrio gigas Characterization and spectroscopic studies
spellingShingle The active centers of adenylylsulfate reductase from Desulfovibrio gigas Characterization and spectroscopic studies
Lampreia, Jorge
Biochemistry
title_short The active centers of adenylylsulfate reductase from Desulfovibrio gigas Characterization and spectroscopic studies
title_full The active centers of adenylylsulfate reductase from Desulfovibrio gigas Characterization and spectroscopic studies
title_fullStr The active centers of adenylylsulfate reductase from Desulfovibrio gigas Characterization and spectroscopic studies
title_full_unstemmed The active centers of adenylylsulfate reductase from Desulfovibrio gigas Characterization and spectroscopic studies
title_sort The active centers of adenylylsulfate reductase from Desulfovibrio gigas Characterization and spectroscopic studies
author Lampreia, Jorge
author_facet Lampreia, Jorge
Moura, Isabel
Teixeira, Miguel
Peck, Harry D.
LeGall, Jean
Huynh, Boi H.
Moura, José J. G.
author_role author
author2 Moura, Isabel
Teixeira, Miguel
Peck, Harry D.
LeGall, Jean
Huynh, Boi H.
Moura, José J. G.
author2_role author
author
author
author
author
author
dc.contributor.none.fl_str_mv Instituto de Tecnologia Química e Biológica António Xavier (ITQB)
RUN
dc.contributor.author.fl_str_mv Lampreia, Jorge
Moura, Isabel
Teixeira, Miguel
Peck, Harry D.
LeGall, Jean
Huynh, Boi H.
Moura, José J. G.
dc.subject.por.fl_str_mv Biochemistry
topic Biochemistry
description NIGMS NIH HHS (GM 032187) NIDDK NIH HHS (DK 01135)
publishDate 1990
dc.date.none.fl_str_mv 1990-01-01
1990-01-01T00:00:00Z
2019-09-10T22:49:51Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv https://doi.org/10.1111/j.1432-1033.1990.tb15447.x
url https://doi.org/10.1111/j.1432-1033.1990.tb15447.x
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 0014-2956
PURE: 14640845
http://www.scopus.com/inward/record.url?scp=0025320295&partnerID=8YFLogxK
https://doi.org/10.1111/j.1432-1033.1990.tb15447.x
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repository.name.fl_str_mv Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
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