Measuring the cytochrome c nitrite reductase activity—practical considerations on the enzyme assays

Detalhes bibliográficos
Autor(a) principal: Silveira, Célia M.
Data de Publicação: 2010
Outros Autores: Besson, Stéphane, Moura, Isabel, Moura, José J. G., Almeida, Maria Gabriela
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositórios Científicos de Acesso Aberto de Portugal (RCAAP)
Texto Completo: http://hdl.handle.net/10362/6970
Resumo: Hindawi Publishing Corporation Bioinorganic Chemistry and Applications Volume 2010, Article ID 634597, 8 pages
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spelling Measuring the cytochrome c nitrite reductase activity—practical considerations on the enzyme assaysHindawi Publishing Corporation Bioinorganic Chemistry and Applications Volume 2010, Article ID 634597, 8 pagesThe cytochrome c nitrite reductase (ccNiR) from Desulfovibrio desulfuricans ATCC 27774 is able to reduce nitrite to ammonia in a six-electron transfer reaction. Although extensively characterized from the spectroscopic and structural points-of-view, some of its kinetic aspects are still under explored. In this work the kinetic behaviour of ccNiR has been evaluated in a systematic manner using two different spectrophotometric assays carried out in the presence of different redox mediators and a direct electrochemical approach. Solution assays have proved that the specific activity of ccNiR decreases with the reduction potential of the electronic carriers and ammonium is always the main product of nitrite reduction. The catalytic parameters were discussed on the basis of the mediator reducing power and also taking into account the location of their putative docking sites with ccNiR. Due to the fast kinetics of ccNiR, electron delivering from reduced electron donors is rate-limiting in all spectrophotometric assays, so the estimated kinetic constants are apparent only. Nevertheless, this limitation could be overcome by using a direct electrochemical approach which shows that the binding affinity for nitrite decreases whilst turnover increases with the reductive driving force.Hindawi Publishing CorporationRUNSilveira, Célia M.Besson, StéphaneMoura, IsabelMoura, José J. G.Almeida, Maria Gabriela2012-02-08T16:35:58Z20102010-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10362/6970eng1565-3633info:eu-repo/semantics/openAccessreponame:Repositórios Científicos de Acesso Aberto de Portugal (RCAAP)instname:FCCN, serviços digitais da FCT – Fundação para a Ciência e a Tecnologiainstacron:RCAAP2024-05-22T17:10:13Zoai:run.unl.pt:10362/6970Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireinfo@rcaap.ptopendoar:https://opendoar.ac.uk/repository/71602025-05-28T16:41:00.523754Repositórios Científicos de Acesso Aberto de Portugal (RCAAP) - FCCN, serviços digitais da FCT – Fundação para a Ciência e a Tecnologiafalse
dc.title.none.fl_str_mv Measuring the cytochrome c nitrite reductase activity—practical considerations on the enzyme assays
title Measuring the cytochrome c nitrite reductase activity—practical considerations on the enzyme assays
spellingShingle Measuring the cytochrome c nitrite reductase activity—practical considerations on the enzyme assays
Silveira, Célia M.
title_short Measuring the cytochrome c nitrite reductase activity—practical considerations on the enzyme assays
title_full Measuring the cytochrome c nitrite reductase activity—practical considerations on the enzyme assays
title_fullStr Measuring the cytochrome c nitrite reductase activity—practical considerations on the enzyme assays
title_full_unstemmed Measuring the cytochrome c nitrite reductase activity—practical considerations on the enzyme assays
title_sort Measuring the cytochrome c nitrite reductase activity—practical considerations on the enzyme assays
author Silveira, Célia M.
author_facet Silveira, Célia M.
Besson, Stéphane
Moura, Isabel
Moura, José J. G.
Almeida, Maria Gabriela
author_role author
author2 Besson, Stéphane
Moura, Isabel
Moura, José J. G.
Almeida, Maria Gabriela
author2_role author
author
author
author
dc.contributor.none.fl_str_mv RUN
dc.contributor.author.fl_str_mv Silveira, Célia M.
Besson, Stéphane
Moura, Isabel
Moura, José J. G.
Almeida, Maria Gabriela
description Hindawi Publishing Corporation Bioinorganic Chemistry and Applications Volume 2010, Article ID 634597, 8 pages
publishDate 2010
dc.date.none.fl_str_mv 2010
2010-01-01T00:00:00Z
2012-02-08T16:35:58Z
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/10362/6970
url http://hdl.handle.net/10362/6970
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 1565-3633
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
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dc.publisher.none.fl_str_mv Hindawi Publishing Corporation
publisher.none.fl_str_mv Hindawi Publishing Corporation
dc.source.none.fl_str_mv reponame:Repositórios Científicos de Acesso Aberto de Portugal (RCAAP)
instname:FCCN, serviços digitais da FCT – Fundação para a Ciência e a Tecnologia
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instname_str FCCN, serviços digitais da FCT – Fundação para a Ciência e a Tecnologia
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reponame_str Repositórios Científicos de Acesso Aberto de Portugal (RCAAP)
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