Publicação
The isolation and characterization of cytochrome c nitrite reductase subunits (NrfA and NrfH) from Desulfovibrio desulfuricans ATCC 27774. Re-evaluation of the spectroscopic data and redox properties
| Resumo: | The cytochrome c nitrite reductase is isolated from the membranes of the sulfate-reducing bacterium Desulfovibrio desulfuricans ATCC 27774 as a heterooligomeric complex composed by two subunits (61 kDa and 19 kDa) containing c-type hemes, encoded by the genes nrfA and nrfH, respectively. The extracted complex has in average a. 2NrfA:1NrfH composition. The separation of ccNiR subunits from one another is accomplished by gel filtration chromatography in the presence of SDS. The amino-acid sequence and biochemical subunits characterization show that NrfA contains five hemes and NrfH four hemes. These considerations enabled the revision of a vast amount of existing spectroscopic data on the NrfHA complex that was not originally well interpreted due to the lack of knowledge on the heme content and the oligomeric enzyme status. Based on EPR and Mössbauer parameters and their correlation to structural information recently obtained from X-ray crystallography on the NrfA structure [Cunha, C.A., Macieira, S., Dias, J.M., Almeida, M.G., Gonçalves, L.M.L., Costa, C., Lampreia, J., Huber, R., Moura, J.J.G., Moura, I. & Romão, M. (2003) J. Biol. Chem. 278, 1745517465], we propose the full assignment of midpoint reduction potentials values to the individual hemes. NrfA contains the high-spin catalytic site (-80 mV) as well as a quite unusual high reduction potential (+150 mV)/low-spin bis-His coordinated heme, considered to be the site where electrons enter. In addition, the reassessment of the spectroscopic data allowed the first partial spectroscopic characterization of the NrfH subunit. The four NrfH hemes are all in a low-spin state (S = 1/2). One of them has a gmax at 3.55, characteristic of bis-histidinyl iron ligands in a noncoplanar arrangement, and has a positive reduction potential. |
|---|---|
| Autores principais: | Almeida, Maria Gabriela |
| Outros Autores: | Macieira, Sofia; Gonçalves, Luisa L.; Huber, Robert; Cunha, Carlos A.; Romão, Maria João; Costa, Cristina; Lampreia, Jorge; Moura, José J. G.; Moura, Isabel |
| Assunto: | c-type hemes EPR Mössbauer Nitrite reductase subunits Redox potentials Biochemistry |
| Ano: | 2003 |
| País: | Portugal |
| Tipo de documento: | artigo |
| Tipo de acesso: | acesso aberto |
| Instituição associada: | Universidade Nova de Lisboa |
| Idioma: | inglês |
| Origem: | Repositório Institucional da UNL |
| _version_ | 1868983327458852864 |
|---|---|
| author | Almeida, Maria Gabriela |
| author2 | Macieira, Sofia Gonçalves, Luisa L. Huber, Robert Cunha, Carlos A. Romão, Maria João Costa, Cristina Lampreia, Jorge Moura, José J. G. Moura, Isabel |
| author2_role | author author author author author author author author author |
| author_facet | Almeida, Maria Gabriela Macieira, Sofia Gonçalves, Luisa L. Huber, Robert Cunha, Carlos A. Romão, Maria João Costa, Cristina Lampreia, Jorge Moura, José J. G. Moura, Isabel |
| author_role | author |
| contributor_name_str_mv | DQ - Departamento de Química CQFB-REQUIMTE - Centro de Química Fina e Biotecnologia (Lab. Associado REQUIMTE) Wiley-Blackwell RUN |
| country_str | PT |
| creators_json_txt | [{\"Person.name\":\"Almeida, Maria Gabriela\"},{\"Person.name\":\"Macieira, Sofia\"},{\"Person.name\":\"Gonçalves, Luisa L.\"},{\"Person.name\":\"Huber, Robert\"},{\"Person.name\":\"Cunha, Carlos A.\"},{\"Person.name\":\"Romão, Maria João\"},{\"Person.name\":\"Costa, Cristina\"},{\"Person.name\":\"Lampreia, Jorge\"},{\"Person.name\":\"Moura, José J. G.\"},{\"Person.name\":\"Moura, Isabel\"}] |
| datacite.contributors.contributor.contributorName.fl_str_mv | DQ - Departamento de Química CQFB-REQUIMTE - Centro de Química Fina e Biotecnologia (Lab. Associado REQUIMTE) Wiley-Blackwell RUN |
| datacite.creators.creator.creatorName.fl_str_mv | Almeida, Maria Gabriela Macieira, Sofia Gonçalves, Luisa L. Huber, Robert Cunha, Carlos A. Romão, Maria João Costa, Cristina Lampreia, Jorge Moura, José J. G. Moura, Isabel |
| datacite.date.Accepted.fl_str_mv | 2003-10-01T00:00:00Z |
| datacite.date.available.fl_str_mv | 2019-03-14T23:15:32Z |
| datacite.date.embargoed.fl_str_mv | 2019-03-14T23:15:32Z |
| datacite.rights.fl_str_mv | http://purl.org/coar/access_right/c_abf2 |
| datacite.subjects.subject.fl_str_mv | c-type hemes EPR Mössbauer Nitrite reductase subunits Redox potentials Biochemistry |
| datacite.titles.title.fl_str_mv | The isolation and characterization of cytochrome c nitrite reductase subunits (NrfA and NrfH) from Desulfovibrio desulfuricans ATCC 27774. Re-evaluation of the spectroscopic data and redox properties |
| dc.contributor.none.fl_str_mv | DQ - Departamento de Química CQFB-REQUIMTE - Centro de Química Fina e Biotecnologia (Lab. Associado REQUIMTE) Wiley-Blackwell RUN |
| dc.creator.none.fl_str_mv | Almeida, Maria Gabriela Macieira, Sofia Gonçalves, Luisa L. Huber, Robert Cunha, Carlos A. Romão, Maria João Costa, Cristina Lampreia, Jorge Moura, José J. G. Moura, Isabel |
| dc.date.Accepted.fl_str_mv | 2003-10-01T00:00:00Z |
| dc.date.available.fl_str_mv | 2019-03-14T23:15:32Z |
| dc.date.embargoed.fl_str_mv | 2019-03-14T23:15:32Z |
| dc.format.none.fl_str_mv | application/pdf |
| dc.identifier.none.fl_str_mv | http://www.scopus.com/inward/record.url?scp=0141704108&partnerID=8YFLogxK |
| dc.language.none.fl_str_mv | eng |
| dc.rights.none.fl_str_mv | http://purl.org/coar/access_right/c_abf2 |
| dc.subject.none.fl_str_mv | c-type hemes EPR Mössbauer Nitrite reductase subunits Redox potentials Biochemistry |
| dc.title.fl_str_mv | The isolation and characterization of cytochrome c nitrite reductase subunits (NrfA and NrfH) from Desulfovibrio desulfuricans ATCC 27774. Re-evaluation of the spectroscopic data and redox properties |
| dc.type.none.fl_str_mv | http://purl.org/coar/resource_type/c_6501 |
| description | The cytochrome c nitrite reductase is isolated from the membranes of the sulfate-reducing bacterium Desulfovibrio desulfuricans ATCC 27774 as a heterooligomeric complex composed by two subunits (61 kDa and 19 kDa) containing c-type hemes, encoded by the genes nrfA and nrfH, respectively. The extracted complex has in average a. 2NrfA:1NrfH composition. The separation of ccNiR subunits from one another is accomplished by gel filtration chromatography in the presence of SDS. The amino-acid sequence and biochemical subunits characterization show that NrfA contains five hemes and NrfH four hemes. These considerations enabled the revision of a vast amount of existing spectroscopic data on the NrfHA complex that was not originally well interpreted due to the lack of knowledge on the heme content and the oligomeric enzyme status. Based on EPR and Mössbauer parameters and their correlation to structural information recently obtained from X-ray crystallography on the NrfA structure [Cunha, C.A., Macieira, S., Dias, J.M., Almeida, M.G., Gonçalves, L.M.L., Costa, C., Lampreia, J., Huber, R., Moura, J.J.G., Moura, I. & Romão, M. (2003) J. Biol. Chem. 278, 1745517465], we propose the full assignment of midpoint reduction potentials values to the individual hemes. NrfA contains the high-spin catalytic site (-80 mV) as well as a quite unusual high reduction potential (+150 mV)/low-spin bis-His coordinated heme, considered to be the site where electrons enter. In addition, the reassessment of the spectroscopic data allowed the first partial spectroscopic characterization of the NrfH subunit. The four NrfH hemes are all in a low-spin state (S = 1/2). One of them has a gmax at 3.55, characteristic of bis-histidinyl iron ligands in a noncoplanar arrangement, and has a positive reduction potential. |
| dirty | 0 |
| eu_rights_str_mv | openAccess |
| format | article |
| fulltext.url.fl_str_mv | https://run.unl.pt/bitstreams/3e8a4025-968d-4bf5-84ce-68b8dc627904/download |
| id | run_b13dcd4bbd2412ec0f05fd0fc4f8a922 |
| identifier.url.fl_str_mv | http://www.scopus.com/inward/record.url?scp=0141704108&partnerID=8YFLogxK |
| inst_facet_str | urn:organizationAcronym:unl{{{_:::_}}}Universidade Nova de Lisboa |
| instacron_str | unl |
| institution | Universidade Nova de Lisboa |
| instname_str | Universidade Nova de Lisboa |
| language | eng |
| network_acronym_str | run |
| network_name_str | Repositório Institucional da UNL |
| oai_identifier_str | oai:run.unl.pt:10362/63331 |
| organization_str_mv | urn:organizationAcronym:unl |
| person_str_mv | Almeida, Maria Gabriela Macieira, Sofia Gonçalves, Luisa L. Huber, Robert Cunha, Carlos A. Romão, Maria João Costa, Cristina Lampreia, Jorge Moura, José J. G. Moura, Isabel |
| publishDate | 2003 |
| repo_facet_str | urn:repositoryAcronym:run{{{_:::_}}}Repositório Institucional da UNL |
| reponame_str | Repositório Institucional da UNL |
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| spelling | engenThe cytochrome c nitrite reductase is isolated from the membranes of the sulfate-reducing bacterium Desulfovibrio desulfuricans ATCC 27774 as a heterooligomeric complex composed by two subunits (61 kDa and 19 kDa) containing c-type hemes, encoded by the genes nrfA and nrfH, respectively. The extracted complex has in average a. 2NrfA:1NrfH composition. The separation of ccNiR subunits from one another is accomplished by gel filtration chromatography in the presence of SDS. The amino-acid sequence and biochemical subunits characterization show that NrfA contains five hemes and NrfH four hemes. These considerations enabled the revision of a vast amount of existing spectroscopic data on the NrfHA complex that was not originally well interpreted due to the lack of knowledge on the heme content and the oligomeric enzyme status. Based on EPR and Mössbauer parameters and their correlation to structural information recently obtained from X-ray crystallography on the NrfA structure [Cunha, C.A., Macieira, S., Dias, J.M., Almeida, M.G., Gonçalves, L.M.L., Costa, C., Lampreia, J., Huber, R., Moura, J.J.G., Moura, I. & Romão, M. (2003) J. Biol. Chem. 278, 1745517465], we propose the full assignment of midpoint reduction potentials values to the individual hemes. NrfA contains the high-spin catalytic site (-80 mV) as well as a quite unusual high reduction potential (+150 mV)/low-spin bis-His coordinated heme, considered to be the site where electrons enter. In addition, the reassessment of the spectroscopic data allowed the first partial spectroscopic characterization of the NrfH subunit. The four NrfH hemes are all in a low-spin state (S = 1/2). One of them has a gmax at 3.55, characteristic of bis-histidinyl iron ligands in a noncoplanar arrangement, and has a positive reduction potential.application/pdfenThe isolation and characterization of cytochrome c nitrite reductase subunits (NrfA and NrfH) from Desulfovibrio desulfuricans ATCC 27774. Re-evaluation of the spectroscopic data and redox propertiesAlmeida, Maria GabrielaMacieira, SofiaGonçalves, Luisa L.Huber, RobertCunha, Carlos A.Romão, Maria JoãoCosta, CristinaLampreia, JorgeMoura, José J. G.Moura, IsabelDQ - Departamento de QuímicaCQFB-REQUIMTE - Centro de Química Fina e Biotecnologia (Lab. Associado REQUIMTE)Wiley-BlackwellHostingInstitutionOrganizationalRUNe-mailmailto:run@unl.ptrun@unl.ptISSNIsPartOf0014-2956URNIsPartOfPURE: 12112597URNIsPartOfPURE UUID: de141574-7dfd-4a6a-9272-e04c0f132062URNIsPartOfScopus: 0141704108URNIsPartOfPubMed: 14511372URNIsPartOfWOS: 000185370100005URNIsPartOfORCID: /0000-0002-3004-0543/work/55386318URNIsPartOfORCID: /0000-0002-8611-9023/work/62320646URNIsPartOfORCID: /0000-0002-4726-2388/work/68772145DOIIsPartOf10.1046/j.1432-1033.2003.03772.x2019-03-14T23:15:32Z2003-10-012003-10-01T00:00:00ZURLhttp://www.scopus.com/inward/record.url?scp=0141704108&partnerID=8YFLogxKhttp://purl.org/coar/access_right/c_abf2open accessc-type hemesEPRMössbauerNitrite reductase subunitsRedox potentialsBiochemistry359507 bytesliteraturehttp://purl.org/coar/resource_type/c_6501journal articlehttp://purl.org/coar/access_right/c_abf2application/pdffulltexthttps://run.unl.pt/bitstreams/3e8a4025-968d-4bf5-84ce-68b8dc627904/download |
| spellingShingle | The isolation and characterization of cytochrome c nitrite reductase subunits (NrfA and NrfH) from Desulfovibrio desulfuricans ATCC 27774. Re-evaluation of the spectroscopic data and redox properties Almeida, Maria Gabriela c-type hemes EPR Mössbauer Nitrite reductase subunits Redox potentials Biochemistry |
| status | SINGLETON |
| subject.fl_str_mv | c-type hemes EPR Mössbauer Nitrite reductase subunits Redox potentials Biochemistry |
| title | The isolation and characterization of cytochrome c nitrite reductase subunits (NrfA and NrfH) from Desulfovibrio desulfuricans ATCC 27774. Re-evaluation of the spectroscopic data and redox properties |
| title_full | The isolation and characterization of cytochrome c nitrite reductase subunits (NrfA and NrfH) from Desulfovibrio desulfuricans ATCC 27774. Re-evaluation of the spectroscopic data and redox properties |
| title_fullStr | The isolation and characterization of cytochrome c nitrite reductase subunits (NrfA and NrfH) from Desulfovibrio desulfuricans ATCC 27774. Re-evaluation of the spectroscopic data and redox properties |
| title_full_unstemmed | The isolation and characterization of cytochrome c nitrite reductase subunits (NrfA and NrfH) from Desulfovibrio desulfuricans ATCC 27774. Re-evaluation of the spectroscopic data and redox properties |
| title_short | The isolation and characterization of cytochrome c nitrite reductase subunits (NrfA and NrfH) from Desulfovibrio desulfuricans ATCC 27774. Re-evaluation of the spectroscopic data and redox properties |
| title_sort | The isolation and characterization of cytochrome c nitrite reductase subunits (NrfA and NrfH) from Desulfovibrio desulfuricans ATCC 27774. Re-evaluation of the spectroscopic data and redox properties |
| topic | c-type hemes EPR Mössbauer Nitrite reductase subunits Redox potentials Biochemistry |
| topic_facet | c-type hemes EPR Mössbauer Nitrite reductase subunits Redox potentials Biochemistry |
| url | http://www.scopus.com/inward/record.url?scp=0141704108&partnerID=8YFLogxK |
| visible | 1 |