Functional EGF domain of the human neuregulin 1α produced in Escherichia coli with accurate disulfide bonds

Detalhes bibliográficos
Autor(a) principal: Ferreira, Arthur Schveitzer
Data de Publicação: 2022
Outros Autores: Lopacinski, Amanda, Batista, Michel, Hiraiwa, Priscila Mazzocchi, Bueno, Natalia Fernanda, Guimarães, Beatriz Gomes, Zanchin, Nilson Ivo Tonin
Tipo de documento: Artigo
Idioma: por
Título da fonte: Repositório Institucional da FIOCRUZ (ARCA)
Texto Completo: https://www.arca.fiocruz.br/handle/icict/56149
Resumo: Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Biologia Estrutural e Engenharia de Proteínas. Curitiba, PR, Brasil. / Universidade Federal do Paraná. Programa de Pós-Graduação em Biologia Celular e Molecular. Curitiba, PR, Brasil.
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spelling Ferreira, Arthur SchveitzerLopacinski, AmandaBatista, MichelHiraiwa, Priscila MazzocchiBueno, Natalia FernandaGuimarães, Beatriz GomesZanchin, Nilson Ivo Tonin2022-12-21T19:52:24Z2022-12-21T19:52:24Z2022FERREIRA, Arthur Schveitzer et al. Functional EGF domain of the human neuregulin 1α produced in Escherichia coli with accurate disulfide bonds. Molecular Biology Reports, p. 1–13, 2022.1573-4978https://www.arca.fiocruz.br/handle/icict/56149porSpringerFunctional EGF domain of the human neuregulin 1α produced in Escherichia coli with accurate disulfide bondsinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleFundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Biologia Estrutural e Engenharia de Proteínas. Curitiba, PR, Brasil. / Universidade Federal do Paraná. Programa de Pós-Graduação em Biologia Celular e Molecular. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Biologia Estrutural e Engenharia de Proteínas. Curitiba, PR, Brasil. / Universidade Federal do Paraná. Programa de Pós-Graduação em Biologia Celular e Molecular. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Mass Spectrometry Facility. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Biologia Estrutural e Engenharia de Proteínas. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Plataforma de Biologia Estrutural Integrada. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Biologia Estrutural e Engenharia de Proteínas. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Biologia Estrutural e Engenharia de Proteínas. Curitiba, PR, Brasil. / Universidade Federal do Paraná. Programa de Pós-Graduação em Biologia Celular e Molecular. Curitiba, PR, Brasil.Neuregulins comprise a large family of growth factors containing an epidermal growth factor (EGF) domain.NRG1 acts in signaling pathways involved in proliferation, apoptosis, migration, diferentiation, and adhesion of manynormal cell types and in human diseases. The EGF domain of NRG1 mediates signaling by interaction with members ofthe ErbB family of receptors. Easy access to correctly folded hNRG1α EGF domain can be a valuable tool to investigate its function in diferent cell types. The EGF domain of hNRG1α was produced in Escherichia coli in fusion with TrxA and purifedafter cleavage of TrxA. Conformation and stability analyses were performed by using biophysical methods and the disulfde bonds were mapped by mass spectrometry. The activity of the hNRG1α EGF domain was demonstrated in cell proliferation and migration assays. Approximately 3.3 mg of hNRG1α EGF domain were obtained starting from a 0.5 L of E. coli culture. Correct formation of the three disulfde bonds was demonstrated by mass spectrometry with high accuracy. Heat denaturation assays monitored by circular dichroism and dynamic light scattering revealed that it is a highly stable protein. The recombinant EGF domain of hNRG1α purifed in this work is highly active, inducing cell proliferation at concentration as low as 0.05 ng/mL. It induces also cell migration as demonstrated by a gap closure assay. The EGF domain of hNRG1α was produced in E. coli with the correct disulfde bonds and presented high stimulation of HeLa cell proliferation and NDFH cell migration.Neuregulin-1EGF Family of ProteinsDisulfidesNeurregulina-1Familia de Proteínas EGFDisulfurosNeuréguline-1Protéines de la famille de l'EGFDisulfuresNeuregulina-1Família de Proteínas EGFDissulfetosinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZLICENSElicense.txtlicense.txttext/plain; charset=utf-82991https://www.arca.fiocruz.br/bitstream/icict/56149/1/license.txt5a560609d32a3863062d77ff32785d58MD51ORIGINALFunctionalEGF_Molecular-Biology_2022ok.pdfFunctionalEGF_Molecular-Biology_2022ok.pdfapplication/pdf2725772https://www.arca.fiocruz.br/bitstream/icict/56149/2/FunctionalEGF_Molecular-Biology_2022ok.pdfce15b8f33b561169171e0f79257c3b25MD52icict/561492023-06-19 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dc.title.en_US.fl_str_mv Functional EGF domain of the human neuregulin 1α produced in Escherichia coli with accurate disulfide bonds
title Functional EGF domain of the human neuregulin 1α produced in Escherichia coli with accurate disulfide bonds
spellingShingle Functional EGF domain of the human neuregulin 1α produced in Escherichia coli with accurate disulfide bonds
Ferreira, Arthur Schveitzer
Neuregulin-1
EGF Family of Proteins
Disulfides
Neurregulina-1
Familia de Proteínas EGF
Disulfuros
Neuréguline-1
Protéines de la famille de l'EGF
Disulfures
Neuregulina-1
Família de Proteínas EGF
Dissulfetos
title_short Functional EGF domain of the human neuregulin 1α produced in Escherichia coli with accurate disulfide bonds
title_full Functional EGF domain of the human neuregulin 1α produced in Escherichia coli with accurate disulfide bonds
title_fullStr Functional EGF domain of the human neuregulin 1α produced in Escherichia coli with accurate disulfide bonds
title_full_unstemmed Functional EGF domain of the human neuregulin 1α produced in Escherichia coli with accurate disulfide bonds
title_sort Functional EGF domain of the human neuregulin 1α produced in Escherichia coli with accurate disulfide bonds
author Ferreira, Arthur Schveitzer
author_facet Ferreira, Arthur Schveitzer
Lopacinski, Amanda
Batista, Michel
Hiraiwa, Priscila Mazzocchi
Bueno, Natalia Fernanda
Guimarães, Beatriz Gomes
Zanchin, Nilson Ivo Tonin
author_role author
author2 Lopacinski, Amanda
Batista, Michel
Hiraiwa, Priscila Mazzocchi
Bueno, Natalia Fernanda
Guimarães, Beatriz Gomes
Zanchin, Nilson Ivo Tonin
author2_role author
author
author
author
author
author
dc.contributor.author.fl_str_mv Ferreira, Arthur Schveitzer
Lopacinski, Amanda
Batista, Michel
Hiraiwa, Priscila Mazzocchi
Bueno, Natalia Fernanda
Guimarães, Beatriz Gomes
Zanchin, Nilson Ivo Tonin
dc.subject.en.en_US.fl_str_mv Neuregulin-1
EGF Family of Proteins
Disulfides
topic Neuregulin-1
EGF Family of Proteins
Disulfides
Neurregulina-1
Familia de Proteínas EGF
Disulfuros
Neuréguline-1
Protéines de la famille de l'EGF
Disulfures
Neuregulina-1
Família de Proteínas EGF
Dissulfetos
dc.subject.es.en_US.fl_str_mv Neurregulina-1
Familia de Proteínas EGF
Disulfuros
dc.subject.fr.en_US.fl_str_mv Neuréguline-1
Protéines de la famille de l'EGF
Disulfures
dc.subject.decs.en_US.fl_str_mv Neuregulina-1
Família de Proteínas EGF
Dissulfetos
description Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Biologia Estrutural e Engenharia de Proteínas. Curitiba, PR, Brasil. / Universidade Federal do Paraná. Programa de Pós-Graduação em Biologia Celular e Molecular. Curitiba, PR, Brasil.
publishDate 2022
dc.date.accessioned.fl_str_mv 2022-12-21T19:52:24Z
dc.date.available.fl_str_mv 2022-12-21T19:52:24Z
dc.date.issued.fl_str_mv 2022
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.citation.fl_str_mv FERREIRA, Arthur Schveitzer et al. Functional EGF domain of the human neuregulin 1α produced in Escherichia coli with accurate disulfide bonds. Molecular Biology Reports, p. 1–13, 2022.
dc.identifier.uri.fl_str_mv https://www.arca.fiocruz.br/handle/icict/56149
dc.identifier.issn.en_US.fl_str_mv 1573-4978
identifier_str_mv FERREIRA, Arthur Schveitzer et al. Functional EGF domain of the human neuregulin 1α produced in Escherichia coli with accurate disulfide bonds. Molecular Biology Reports, p. 1–13, 2022.
1573-4978
url https://www.arca.fiocruz.br/handle/icict/56149
dc.language.iso.fl_str_mv por
language por
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Springer
publisher.none.fl_str_mv Springer
dc.source.none.fl_str_mv reponame:Repositório Institucional da FIOCRUZ (ARCA)
instname:Fundação Oswaldo Cruz (FIOCRUZ)
instacron:FIOCRUZ
instname_str Fundação Oswaldo Cruz (FIOCRUZ)
instacron_str FIOCRUZ
institution FIOCRUZ
reponame_str Repositório Institucional da FIOCRUZ (ARCA)
collection Repositório Institucional da FIOCRUZ (ARCA)
bitstream.url.fl_str_mv https://www.arca.fiocruz.br/bitstream/icict/56149/1/license.txt
https://www.arca.fiocruz.br/bitstream/icict/56149/2/FunctionalEGF_Molecular-Biology_2022ok.pdf
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repository.name.fl_str_mv Repositório Institucional da FIOCRUZ (ARCA) - Fundação Oswaldo Cruz (FIOCRUZ)
repository.mail.fl_str_mv repositorio.arca@fiocruz.br
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