Camelid nanobodies raised against an integral membrane enzyme, nitric oxide reductase

Detalhes bibliográficos
Autor(a) principal: Conrath, Katja
Data de Publicação: 2009
Outros Autores: Pereira, Alice S., Martins, Carlos E., Timóteo, Cristina G., Tavares, Pedro, Moura, Isabel, Moura, José J. G.
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10362/8693
Resumo: Protein Sci. 2009 Mar;18(3):619-28. doi: 10.1002/pro.69.
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spelling Camelid nanobodies raised against an integral membrane enzyme, nitric oxide reductaseNitric oxide reductaseCamelid antibodiesVHH domainSPRPhage displayProtein Sci. 2009 Mar;18(3):619-28. doi: 10.1002/pro.69.Nitric Oxide Reductase (NOR) is an integral membrane protein performing the reduction of NO to N2O. NOR is composed of two subunits: the large one (NorB) is a bundle of 12 transmembrane helices (TMH). It contains a b type heme and a binuclear iron site, which is believed to be the catalytic site, comprising a heme b and a non-hemic iron. The small subunit(NorC) harbors a cytochrome c and is attached to the membrane through a unique TMH. With the aim to perform structural and functional studies of NOR, we have immunized dromedaries with NOR and produced several antibody fragments of the heavy chain (VHHs, also known as nanobodiesTM). These fragments have been used to develop a faster NOR purification procedure, to proceed to crystallization assays and to analyze the electron transfer of electron donors. BIAcore experiments have revealed that up to three VHHs can bind concomitantly to NOR with affinities in the nanomolar range. This is the first example of the use of VHHs with an integral membrane protein. Our results indicate that VHHs are able to recognize with high affinity distinct epitopes on this class of proteins, and can be used as versatile and valuable tool for purification, functional study and crystallization of integral membrane proteins.Wiley-BlackwellRUNConrath, KatjaPereira, Alice S.Martins, Carlos E.Timóteo, Cristina G.Tavares, PedroMoura, IsabelMoura, José J. G.2013-02-05T12:10:59Z20092009-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10362/8693eng0961-8368info: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:RCAAP2024-03-11T03:41:34Zoai:run.unl.pt:10362/8693Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T03:18:22.703939Repositório 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 Camelid nanobodies raised against an integral membrane enzyme, nitric oxide reductase
title Camelid nanobodies raised against an integral membrane enzyme, nitric oxide reductase
spellingShingle Camelid nanobodies raised against an integral membrane enzyme, nitric oxide reductase
Conrath, Katja
Nitric oxide reductase
Camelid antibodies
VHH domain
SPR
Phage display
title_short Camelid nanobodies raised against an integral membrane enzyme, nitric oxide reductase
title_full Camelid nanobodies raised against an integral membrane enzyme, nitric oxide reductase
title_fullStr Camelid nanobodies raised against an integral membrane enzyme, nitric oxide reductase
title_full_unstemmed Camelid nanobodies raised against an integral membrane enzyme, nitric oxide reductase
title_sort Camelid nanobodies raised against an integral membrane enzyme, nitric oxide reductase
author Conrath, Katja
author_facet Conrath, Katja
Pereira, Alice S.
Martins, Carlos E.
Timóteo, Cristina G.
Tavares, Pedro
Moura, Isabel
Moura, José J. G.
author_role author
author2 Pereira, Alice S.
Martins, Carlos E.
Timóteo, Cristina G.
Tavares, Pedro
Moura, Isabel
Moura, José J. G.
author2_role author
author
author
author
author
author
dc.contributor.none.fl_str_mv RUN
dc.contributor.author.fl_str_mv Conrath, Katja
Pereira, Alice S.
Martins, Carlos E.
Timóteo, Cristina G.
Tavares, Pedro
Moura, Isabel
Moura, José J. G.
dc.subject.por.fl_str_mv Nitric oxide reductase
Camelid antibodies
VHH domain
SPR
Phage display
topic Nitric oxide reductase
Camelid antibodies
VHH domain
SPR
Phage display
description Protein Sci. 2009 Mar;18(3):619-28. doi: 10.1002/pro.69.
publishDate 2009
dc.date.none.fl_str_mv 2009
2009-01-01T00:00:00Z
2013-02-05T12:10:59Z
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/10362/8693
url http://hdl.handle.net/10362/8693
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 0961-8368
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
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dc.publisher.none.fl_str_mv Wiley-Blackwell
publisher.none.fl_str_mv Wiley-Blackwell
dc.source.none.fl_str_mv reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
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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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