Intact protein folding in the glutathione-depleted endoplasmic reticulum implicates alternative protein thiol reductants
Autor(a) principal: | |
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Data de Publicação: | 2014 |
Outros Autores: | , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10400.1/11452 |
Resumo: | Protein folding homeostasis in the endoplasmic reticulum (ER) requires efficient protein thiol oxidation, but also relies on a parallel reductive process to edit disulfides during the maturation or degradation of secreted proteins. To critically examine the widely held assumption that reduced ER glutathione fuels disulfide reduction, we expressed a modified form of a cytosolic glutathione-degrading enzyme, ChaC1, in the ER lumen. ChaC1(CtoS) purged the ER of glutathione eliciting the expected kinetic defect in oxidation of an ER-localized glutathione-coupled Grx1-roGFP2 optical probe, but had no effect on the disulfide editing-dependent maturation of the LDL receptor or the reduction-dependent degradation of misfolded alpha-1 antitrypsin. Furthermore, glutathione depletion had no measurable effect on induction of the unfolded protein response (UPR); a sensitive measure of ER protein folding homeostasis. These findings challenge the importance of reduced ER glutathione and suggest the existence of alternative electron donor(s) that maintain the reductive capacity of the ER. |
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Intact protein folding in the glutathione-depleted endoplasmic reticulum implicates alternative protein thiol reductantsDisulfide-bond formationLocalized peroxiredoxinOxidized glutathioneReducing pathwaysRedoxIsomeraseErCatalysisOxidationErdj5Protein folding homeostasis in the endoplasmic reticulum (ER) requires efficient protein thiol oxidation, but also relies on a parallel reductive process to edit disulfides during the maturation or degradation of secreted proteins. To critically examine the widely held assumption that reduced ER glutathione fuels disulfide reduction, we expressed a modified form of a cytosolic glutathione-degrading enzyme, ChaC1, in the ER lumen. ChaC1(CtoS) purged the ER of glutathione eliciting the expected kinetic defect in oxidation of an ER-localized glutathione-coupled Grx1-roGFP2 optical probe, but had no effect on the disulfide editing-dependent maturation of the LDL receptor or the reduction-dependent degradation of misfolded alpha-1 antitrypsin. Furthermore, glutathione depletion had no measurable effect on induction of the unfolded protein response (UPR); a sensitive measure of ER protein folding homeostasis. These findings challenge the importance of reduced ER glutathione and suggest the existence of alternative electron donor(s) that maintain the reductive capacity of the ER.Wellcome Trust [084812/Z/08/Z, 100140]; European Commission EU [277713]; Fundacao para a Ciencia e a Tecnologia [PTDC/QUI-BIQ/119677/2010]Elife Sciences Publications LtdSapientiaTsunoda, SatoshiAvezov, EdwardZyryanova, AlisaKonno, TasukuLeonardo Mendes-SilvaMelo, EduardoHarding, Heather P.Ron, David2018-12-07T14:53:19Z2014-072014-07-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10400.1/11452eng2050-084X10.7554/eLife.03421info: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:RCAAP2023-07-24T10:23:15Zoai:sapientia.ualg.pt:10400.1/11452Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T20:02:57.547063Repositório 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 |
Intact protein folding in the glutathione-depleted endoplasmic reticulum implicates alternative protein thiol reductants |
title |
Intact protein folding in the glutathione-depleted endoplasmic reticulum implicates alternative protein thiol reductants |
spellingShingle |
Intact protein folding in the glutathione-depleted endoplasmic reticulum implicates alternative protein thiol reductants Tsunoda, Satoshi Disulfide-bond formation Localized peroxiredoxin Oxidized glutathione Reducing pathways Redox Isomerase Er Catalysis Oxidation Erdj5 |
title_short |
Intact protein folding in the glutathione-depleted endoplasmic reticulum implicates alternative protein thiol reductants |
title_full |
Intact protein folding in the glutathione-depleted endoplasmic reticulum implicates alternative protein thiol reductants |
title_fullStr |
Intact protein folding in the glutathione-depleted endoplasmic reticulum implicates alternative protein thiol reductants |
title_full_unstemmed |
Intact protein folding in the glutathione-depleted endoplasmic reticulum implicates alternative protein thiol reductants |
title_sort |
Intact protein folding in the glutathione-depleted endoplasmic reticulum implicates alternative protein thiol reductants |
author |
Tsunoda, Satoshi |
author_facet |
Tsunoda, Satoshi Avezov, Edward Zyryanova, Alisa Konno, Tasuku Leonardo Mendes-Silva Melo, Eduardo Harding, Heather P. Ron, David |
author_role |
author |
author2 |
Avezov, Edward Zyryanova, Alisa Konno, Tasuku Leonardo Mendes-Silva Melo, Eduardo Harding, Heather P. Ron, David |
author2_role |
author author author author author author author |
dc.contributor.none.fl_str_mv |
Sapientia |
dc.contributor.author.fl_str_mv |
Tsunoda, Satoshi Avezov, Edward Zyryanova, Alisa Konno, Tasuku Leonardo Mendes-Silva Melo, Eduardo Harding, Heather P. Ron, David |
dc.subject.por.fl_str_mv |
Disulfide-bond formation Localized peroxiredoxin Oxidized glutathione Reducing pathways Redox Isomerase Er Catalysis Oxidation Erdj5 |
topic |
Disulfide-bond formation Localized peroxiredoxin Oxidized glutathione Reducing pathways Redox Isomerase Er Catalysis Oxidation Erdj5 |
description |
Protein folding homeostasis in the endoplasmic reticulum (ER) requires efficient protein thiol oxidation, but also relies on a parallel reductive process to edit disulfides during the maturation or degradation of secreted proteins. To critically examine the widely held assumption that reduced ER glutathione fuels disulfide reduction, we expressed a modified form of a cytosolic glutathione-degrading enzyme, ChaC1, in the ER lumen. ChaC1(CtoS) purged the ER of glutathione eliciting the expected kinetic defect in oxidation of an ER-localized glutathione-coupled Grx1-roGFP2 optical probe, but had no effect on the disulfide editing-dependent maturation of the LDL receptor or the reduction-dependent degradation of misfolded alpha-1 antitrypsin. Furthermore, glutathione depletion had no measurable effect on induction of the unfolded protein response (UPR); a sensitive measure of ER protein folding homeostasis. These findings challenge the importance of reduced ER glutathione and suggest the existence of alternative electron donor(s) that maintain the reductive capacity of the ER. |
publishDate |
2014 |
dc.date.none.fl_str_mv |
2014-07 2014-07-01T00:00:00Z 2018-12-07T14:53:19Z |
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/10400.1/11452 |
url |
http://hdl.handle.net/10400.1/11452 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
2050-084X 10.7554/eLife.03421 |
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 |
Elife Sciences Publications Ltd |
publisher.none.fl_str_mv |
Elife Sciences Publications Ltd |
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 |
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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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1799133263520858112 |