Zinc-substituted desulfovibrio gigas desulforedoxins

Detalhes bibliográficos
Autor(a) principal: Moura, Isabel
Data de Publicação: 2002
Outros Autores: Goodfellow, Brian J., Nunes, Sofia G., Rusnak, Frank, Ascenso, Carla, Moura, José J. G., Volkman, Brian F., Markley, John L.
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10362/1655
Resumo: Protein Science (2002), 11:2464–2470
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spelling Zinc-substituted desulfovibrio gigas desulforedoxinsResolving subunit degeneracy with nonsymmetric pseudocontact shiftsNMRPseudocontact shiftsDesulforedoxin[Fe-4S] centerParamagnetic proteinDesulfovibrio gigasProtein Science (2002), 11:2464–2470Desulfovibrio gigas desulforedoxin (Dx) consists of two identical peptides, each containing one [Fe-4S]center per monomer. Variants with different iron and zinc metal compositions arise when desulforedoxin is produced recombinantly from Escherichia coli. The three forms of the protein, the two homodimers [Fe(III)/Fe(III)]Dx and [Zn(II)/Zn(II)]Dx, and the heterodimer [Fe(III)/Zn(II)]Dx, can be separated by ion exchange chromatography on the basis of their charge differences. Once separated, the desulforedoxins containing iron can be reduced with added dithionite. For NMR studies, different protein samples were prepared labeled with 15N or 15N + 13C. Spectral assignments were determined for [Fe(II)/Fe(II)]Dx and [Fe(II)/Zn(II)]Dx from 3D 15N TOCSY-HSQC and NOESY-HSQC data, and compared with those reported previously for [Zn(II)/Zn(II)]Dx. Assignments for the 13C shifts were obtained from an HNCA experiment. Comparison of 1H–15N HSQC spectra of [Zn(II)/Zn(II)]Dx, [Fe(II)/Fe(II)]Dx and [Fe(II)/Zn(II)]Dx revealed that the pseudocontact shifts in [Fe(II)/Zn(II)]Dx can be decomposed into inter- and intramonomer components, which, when summed, accurately predict the observed pseudocontact shifts observed for [Fe(II)/Fe(II)]Dx. The degree of linearity observed in the pseudocontact shifts for residues 8.5 Å from the metal center indicates that the replacement of Fe(II) by Zn(II) produces little or no change in the structure of Dx. The results suggest a general strategy for the analysis of NMR spectra of homo-oligomeric proteins in which a paramagnetic center introduced into a single subunit is used to break the magnetic symmetry and make it possible to obtain distance constraints (both pseudocontact and NOE) between subunits.Cold Spring Harbor Laboratory PressRUNMoura, IsabelGoodfellow, Brian J.Nunes, Sofia G.Rusnak, FrankAscenso, CarlaMoura, José J. G.Volkman, Brian F.Markley, John L.2008-09-23T13:06:21Z20022002-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10362/1655enginfo: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:RCAAP2024-03-11T03:31:38Zoai:run.unl.pt:10362/1655Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T03:14:46.849158Repositório 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 Zinc-substituted desulfovibrio gigas desulforedoxins
Resolving subunit degeneracy with nonsymmetric pseudocontact shifts
title Zinc-substituted desulfovibrio gigas desulforedoxins
spellingShingle Zinc-substituted desulfovibrio gigas desulforedoxins
Moura, Isabel
NMR
Pseudocontact shifts
Desulforedoxin
[Fe-4S] center
Paramagnetic protein
Desulfovibrio gigas
title_short Zinc-substituted desulfovibrio gigas desulforedoxins
title_full Zinc-substituted desulfovibrio gigas desulforedoxins
title_fullStr Zinc-substituted desulfovibrio gigas desulforedoxins
title_full_unstemmed Zinc-substituted desulfovibrio gigas desulforedoxins
title_sort Zinc-substituted desulfovibrio gigas desulforedoxins
author Moura, Isabel
author_facet Moura, Isabel
Goodfellow, Brian J.
Nunes, Sofia G.
Rusnak, Frank
Ascenso, Carla
Moura, José J. G.
Volkman, Brian F.
Markley, John L.
author_role author
author2 Goodfellow, Brian J.
Nunes, Sofia G.
Rusnak, Frank
Ascenso, Carla
Moura, José J. G.
Volkman, Brian F.
Markley, John L.
author2_role author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv RUN
dc.contributor.author.fl_str_mv Moura, Isabel
Goodfellow, Brian J.
Nunes, Sofia G.
Rusnak, Frank
Ascenso, Carla
Moura, José J. G.
Volkman, Brian F.
Markley, John L.
dc.subject.por.fl_str_mv NMR
Pseudocontact shifts
Desulforedoxin
[Fe-4S] center
Paramagnetic protein
Desulfovibrio gigas
topic NMR
Pseudocontact shifts
Desulforedoxin
[Fe-4S] center
Paramagnetic protein
Desulfovibrio gigas
description Protein Science (2002), 11:2464–2470
publishDate 2002
dc.date.none.fl_str_mv 2002
2002-01-01T00:00:00Z
2008-09-23T13:06:21Z
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 http://hdl.handle.net/10362/1655
url http://hdl.handle.net/10362/1655
dc.language.iso.fl_str_mv eng
language eng
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv Cold Spring Harbor Laboratory Press
publisher.none.fl_str_mv Cold Spring Harbor Laboratory Press
dc.source.none.fl_str_mv reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron:RCAAP
instname_str Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron_str RCAAP
institution RCAAP
reponame_str Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
collection Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
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
repository.mail.fl_str_mv
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