Cofactor composition and function of a H 2 -sensing regulatory hydrogenase as revealed by Mössbauer and EPR spectroscopy

Autores
Roncaroli, Federico; Bill, Eckhard; Friedrich, Bärbel; Lenz, Oliver; Lubitz, Wolfgang; Pandelia, Maria Eirini
Año de publicación
2015
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
The regulatory hydrogenase (RH) from Ralstonia eutropha H16 acts as a sensor for the detection of environmental H2 and regulates gene expression related to hydrogenase-mediated cellular metabolism. In marked contrast to prototypical energy-converting [NiFe] hydrogenases, the RH is apparently insensitive to inhibition by O2 and CO. While the physiological function of regulatory hydrogenases is well established, little is known about the redox cycling of the [NiFe] center and the nature of the iron–sulfur (FeS) clusters acting as electron relay. The absence of any FeS cluster signals in EPR had been attributed to their particular nature, whereas the observation of essentially only two active site redox states, namely Ni-SI and Ni-C, invoked a different operant mechanism. In the present work, we employ a combination of Mössbauer, FTIR and EPR spectroscopic techniques to study the RH, and the results are consistent with the presence of three [4Fe–4S] centers in the small subunit. In the as-isolated, oxidized RH all FeS clusters reside in the EPR-silent 2+ state. Incubation with H2 leads to reduction of two of the [4Fe–4S] clusters, whereas only strongly reducing agents lead to reduction of the third cluster, which is ascribed to be the [4Fe–4S] center in ‘proximal’ position to the [NiFe] center. In the two different active site redox states, the low-spin FeII exhibits distinct Mössbauer features attributed to changes in the electronic and geometric structure of the catalytic center. The results are discussed with regard to the spectral characteristics and physiological function of H2-sensing regulatory hydrogenases.
Fil: Roncaroli, Federico. Max Planck Institut für Chemische Energiekonversion; Alemania. Comisión Nacional de Energía Atómica. Centro Atómico Constituyentes; Argentina
Fil: Bill, Eckhard. Max Planck Institut für Chemische Energiekonversion; Alemania
Fil: Friedrich, Bärbel. Universität zu Berlin; Alemania
Fil: Lenz, Oliver. Universität zu Berlin; Alemania. Technische Universität Berlin; Alemania
Fil: Lubitz, Wolfgang. Max Planck Institut für Chemische Energiekonversion; Alemania
Fil: Pandelia, Maria Eirini. Max Planck Institut für Chemische Energiekonversion; Alemania. State University of Pennsylvania; Estados Unidos
Materia
Regulatory Hydrogenase
pulse EPR
Mössbauer
FeS clusters
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/51940

id CONICETDig_2ba65350d6e1617d32dc27cb03f0d828
oai_identifier_str oai:ri.conicet.gov.ar:11336/51940
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Cofactor composition and function of a H 2 -sensing regulatory hydrogenase as revealed by Mössbauer and EPR spectroscopyRoncaroli, FedericoBill, EckhardFriedrich, BärbelLenz, OliverLubitz, WolfgangPandelia, Maria EiriniRegulatory Hydrogenasepulse EPRMössbauerFeS clustershttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1The regulatory hydrogenase (RH) from Ralstonia eutropha H16 acts as a sensor for the detection of environmental H2 and regulates gene expression related to hydrogenase-mediated cellular metabolism. In marked contrast to prototypical energy-converting [NiFe] hydrogenases, the RH is apparently insensitive to inhibition by O2 and CO. While the physiological function of regulatory hydrogenases is well established, little is known about the redox cycling of the [NiFe] center and the nature of the iron–sulfur (FeS) clusters acting as electron relay. The absence of any FeS cluster signals in EPR had been attributed to their particular nature, whereas the observation of essentially only two active site redox states, namely Ni-SI and Ni-C, invoked a different operant mechanism. In the present work, we employ a combination of Mössbauer, FTIR and EPR spectroscopic techniques to study the RH, and the results are consistent with the presence of three [4Fe–4S] centers in the small subunit. In the as-isolated, oxidized RH all FeS clusters reside in the EPR-silent 2+ state. Incubation with H2 leads to reduction of two of the [4Fe–4S] clusters, whereas only strongly reducing agents lead to reduction of the third cluster, which is ascribed to be the [4Fe–4S] center in ‘proximal’ position to the [NiFe] center. In the two different active site redox states, the low-spin FeII exhibits distinct Mössbauer features attributed to changes in the electronic and geometric structure of the catalytic center. The results are discussed with regard to the spectral characteristics and physiological function of H2-sensing regulatory hydrogenases.Fil: Roncaroli, Federico. Max Planck Institut für Chemische Energiekonversion; Alemania. Comisión Nacional de Energía Atómica. Centro Atómico Constituyentes; ArgentinaFil: Bill, Eckhard. Max Planck Institut für Chemische Energiekonversion; AlemaniaFil: Friedrich, Bärbel. Universität zu Berlin; AlemaniaFil: Lenz, Oliver. Universität zu Berlin; Alemania. Technische Universität Berlin; AlemaniaFil: Lubitz, Wolfgang. Max Planck Institut für Chemische Energiekonversion; AlemaniaFil: Pandelia, Maria Eirini. Max Planck Institut für Chemische Energiekonversion; Alemania. State University of Pennsylvania; Estados UnidosRoyal Society of Chemistry2015-05info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/zipapplication/pdfhttp://hdl.handle.net/11336/51940Roncaroli, Federico; Bill, Eckhard; Friedrich, Bärbel; Lenz, Oliver; Lubitz, Wolfgang; et al.; Cofactor composition and function of a H 2 -sensing regulatory hydrogenase as revealed by Mössbauer and EPR spectroscopy; Royal Society of Chemistry; Chemical Science; 6; 8; 5-2015; 4495-45072041-6539CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1039/C5SC01560Jinfo:eu-repo/semantics/altIdentifier/url/http://pubs.rsc.org/en/content/articlelanding/2015/sc/c5sc01560jinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T09:36:31Zoai:ri.conicet.gov.ar:11336/51940instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-29 09:36:32.168CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Cofactor composition and function of a H 2 -sensing regulatory hydrogenase as revealed by Mössbauer and EPR spectroscopy
title Cofactor composition and function of a H 2 -sensing regulatory hydrogenase as revealed by Mössbauer and EPR spectroscopy
spellingShingle Cofactor composition and function of a H 2 -sensing regulatory hydrogenase as revealed by Mössbauer and EPR spectroscopy
Roncaroli, Federico
Regulatory Hydrogenase
pulse EPR
Mössbauer
FeS clusters
title_short Cofactor composition and function of a H 2 -sensing regulatory hydrogenase as revealed by Mössbauer and EPR spectroscopy
title_full Cofactor composition and function of a H 2 -sensing regulatory hydrogenase as revealed by Mössbauer and EPR spectroscopy
title_fullStr Cofactor composition and function of a H 2 -sensing regulatory hydrogenase as revealed by Mössbauer and EPR spectroscopy
title_full_unstemmed Cofactor composition and function of a H 2 -sensing regulatory hydrogenase as revealed by Mössbauer and EPR spectroscopy
title_sort Cofactor composition and function of a H 2 -sensing regulatory hydrogenase as revealed by Mössbauer and EPR spectroscopy
dc.creator.none.fl_str_mv Roncaroli, Federico
Bill, Eckhard
Friedrich, Bärbel
Lenz, Oliver
Lubitz, Wolfgang
Pandelia, Maria Eirini
author Roncaroli, Federico
author_facet Roncaroli, Federico
Bill, Eckhard
Friedrich, Bärbel
Lenz, Oliver
Lubitz, Wolfgang
Pandelia, Maria Eirini
author_role author
author2 Bill, Eckhard
Friedrich, Bärbel
Lenz, Oliver
Lubitz, Wolfgang
Pandelia, Maria Eirini
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv Regulatory Hydrogenase
pulse EPR
Mössbauer
FeS clusters
topic Regulatory Hydrogenase
pulse EPR
Mössbauer
FeS clusters
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv The regulatory hydrogenase (RH) from Ralstonia eutropha H16 acts as a sensor for the detection of environmental H2 and regulates gene expression related to hydrogenase-mediated cellular metabolism. In marked contrast to prototypical energy-converting [NiFe] hydrogenases, the RH is apparently insensitive to inhibition by O2 and CO. While the physiological function of regulatory hydrogenases is well established, little is known about the redox cycling of the [NiFe] center and the nature of the iron–sulfur (FeS) clusters acting as electron relay. The absence of any FeS cluster signals in EPR had been attributed to their particular nature, whereas the observation of essentially only two active site redox states, namely Ni-SI and Ni-C, invoked a different operant mechanism. In the present work, we employ a combination of Mössbauer, FTIR and EPR spectroscopic techniques to study the RH, and the results are consistent with the presence of three [4Fe–4S] centers in the small subunit. In the as-isolated, oxidized RH all FeS clusters reside in the EPR-silent 2+ state. Incubation with H2 leads to reduction of two of the [4Fe–4S] clusters, whereas only strongly reducing agents lead to reduction of the third cluster, which is ascribed to be the [4Fe–4S] center in ‘proximal’ position to the [NiFe] center. In the two different active site redox states, the low-spin FeII exhibits distinct Mössbauer features attributed to changes in the electronic and geometric structure of the catalytic center. The results are discussed with regard to the spectral characteristics and physiological function of H2-sensing regulatory hydrogenases.
Fil: Roncaroli, Federico. Max Planck Institut für Chemische Energiekonversion; Alemania. Comisión Nacional de Energía Atómica. Centro Atómico Constituyentes; Argentina
Fil: Bill, Eckhard. Max Planck Institut für Chemische Energiekonversion; Alemania
Fil: Friedrich, Bärbel. Universität zu Berlin; Alemania
Fil: Lenz, Oliver. Universität zu Berlin; Alemania. Technische Universität Berlin; Alemania
Fil: Lubitz, Wolfgang. Max Planck Institut für Chemische Energiekonversion; Alemania
Fil: Pandelia, Maria Eirini. Max Planck Institut für Chemische Energiekonversion; Alemania. State University of Pennsylvania; Estados Unidos
description The regulatory hydrogenase (RH) from Ralstonia eutropha H16 acts as a sensor for the detection of environmental H2 and regulates gene expression related to hydrogenase-mediated cellular metabolism. In marked contrast to prototypical energy-converting [NiFe] hydrogenases, the RH is apparently insensitive to inhibition by O2 and CO. While the physiological function of regulatory hydrogenases is well established, little is known about the redox cycling of the [NiFe] center and the nature of the iron–sulfur (FeS) clusters acting as electron relay. The absence of any FeS cluster signals in EPR had been attributed to their particular nature, whereas the observation of essentially only two active site redox states, namely Ni-SI and Ni-C, invoked a different operant mechanism. In the present work, we employ a combination of Mössbauer, FTIR and EPR spectroscopic techniques to study the RH, and the results are consistent with the presence of three [4Fe–4S] centers in the small subunit. In the as-isolated, oxidized RH all FeS clusters reside in the EPR-silent 2+ state. Incubation with H2 leads to reduction of two of the [4Fe–4S] clusters, whereas only strongly reducing agents lead to reduction of the third cluster, which is ascribed to be the [4Fe–4S] center in ‘proximal’ position to the [NiFe] center. In the two different active site redox states, the low-spin FeII exhibits distinct Mössbauer features attributed to changes in the electronic and geometric structure of the catalytic center. The results are discussed with regard to the spectral characteristics and physiological function of H2-sensing regulatory hydrogenases.
publishDate 2015
dc.date.none.fl_str_mv 2015-05
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/51940
Roncaroli, Federico; Bill, Eckhard; Friedrich, Bärbel; Lenz, Oliver; Lubitz, Wolfgang; et al.; Cofactor composition and function of a H 2 -sensing regulatory hydrogenase as revealed by Mössbauer and EPR spectroscopy; Royal Society of Chemistry; Chemical Science; 6; 8; 5-2015; 4495-4507
2041-6539
CONICET Digital
CONICET
url http://hdl.handle.net/11336/51940
identifier_str_mv Roncaroli, Federico; Bill, Eckhard; Friedrich, Bärbel; Lenz, Oliver; Lubitz, Wolfgang; et al.; Cofactor composition and function of a H 2 -sensing regulatory hydrogenase as revealed by Mössbauer and EPR spectroscopy; Royal Society of Chemistry; Chemical Science; 6; 8; 5-2015; 4495-4507
2041-6539
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1039/C5SC01560J
info:eu-repo/semantics/altIdentifier/url/http://pubs.rsc.org/en/content/articlelanding/2015/sc/c5sc01560j
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/zip
application/pdf
dc.publisher.none.fl_str_mv Royal Society of Chemistry
publisher.none.fl_str_mv Royal Society of Chemistry
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
_version_ 1844613146428309504
score 13.070432