A toolkit for recombinant production of seven human EGF family growth factors in active conformation

Detalhes bibliográficos
Autor(a) principal: Ferreira, Arthur Schveitzer
Data de Publicação: 2022
Outros Autores: Lopacinski, Amanda, Batista, Michel, Hiraiwa, Priscila Mazzocchi, 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/52522
Resumo: Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Biologia Estrutural e Engenharia de Proteínas. Curitiba, PR, Brasil. / Universidade Federal do Paraná. Departamento de Biologia Celular e Molecular. Curitiba, PR, Brasil.
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spelling Ferreira, Arthur SchveitzerLopacinski, AmandaBatista, MichelHiraiwa, Priscila MazzocchiGuimarães, Beatriz GomesZanchin, Nilson Ivo Tonin2022-05-03T19:42:40Z2022-05-03T19:42:40Z2022FERREIRA, Arthur Schveitzer et al. A toolkit for recombinant production of seven human EGF family growth factors in active conformation. Scientific Reports, v. 12, n. 5034, p. 1–14, 2022.2045-2322https://www.arca.fiocruz.br/handle/icict/5252210.1038/s41598-022-09060-9porNatureEscherichia coliEGF Family of ProteinsBiotechnologyCell ProliferationPathologyFamilia de Proteínas EGFBiotecnologíaProliferación CelularPatologíaProtéines de la famille de l'EGFBiotechnologieProlifération cellulaireAnatomopathologieFamília de Proteínas EGFBiotecnologiaProliferação de CélulasPatologiaA toolkit for recombinant production of seven human EGF family growth factors in active conformationinfo: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. Departamento de 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. Departamento de Biologia Celular e Molecular. Curitiba, PR, Brasil.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Mass Spectrometry Facility - RPT02H. 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.Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Biologia Estrutural e Engenharia de Proteínas. Curitiba, PR, Brasil. / Universidade Federal do Paraná. Departamento de Biologia Celular e Molecular. Curitiba, PR, Brasil.Epidermal growth factors (EGF) play a wide range of roles in embryogenesis, skin development, immune response homeostasis. They are involved in several pathologies as well, including several cancer types, psoriasis, chronic pain and chronic kidney disease. All members share the structural EGF domain, which is responsible for receptor interaction, thereby initiating transduction of signals. EGF growth factors have intense use in fundamental research and high potential for biotechnological applications. However, due to their structural organization with three disulfde bonds, recombinant production of these factors in prokaryotic systems is not straightforward. A signifcant fraction usually forms inclusion bodies. For the fraction remaining soluble, misfolding and incomplete disulfde bond formation may afect the amount of active factor in solution, which can compromise experimental conclusions and biotechnological applications. In this work, we describe a reliable procedure to produce seven human growth factors of the EGF family in Escherichia coli. Biophysical and stability analyses using limited proteolysis, light scattering, circular dichroism and nanoDSF show that the recombinant factors present folded and stable conformation. Cell proliferation and scratch healing assays confrmed that the recombinant factors are highly active at concentrations as low as 5 ng/ml.info:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZLICENSElicense.txtlicense.txttext/plain; charset=utf-83084https://www.arca.fiocruz.br/bitstream/icict/52522/1/license.txt783568c2893d2e25a99990b126be1772MD51ORIGINAL41598_2022_Article_9060OK.pdf41598_2022_Article_9060OK.pdfapplication/pdf5045524https://www.arca.fiocruz.br/bitstream/icict/52522/2/41598_2022_Article_9060OK.pdf51f425eccc4a58c72dfab7479f32e7e4MD52icict/525222022-05-03 16:42:40.883oai:www.arca.fiocruz.br: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ório InstitucionalPUBhttps://www.arca.fiocruz.br/oai/requestrepositorio.arca@fiocruz.bropendoar:21352022-05-03T19:42:40Repositório Institucional da FIOCRUZ (ARCA) - 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dc.title.pt_BR.fl_str_mv A toolkit for recombinant production of seven human EGF family growth factors in active conformation
title A toolkit for recombinant production of seven human EGF family growth factors in active conformation
spellingShingle A toolkit for recombinant production of seven human EGF family growth factors in active conformation
Ferreira, Arthur Schveitzer
Escherichia coli
EGF Family of Proteins
Biotechnology
Cell Proliferation
Pathology
Familia de Proteínas EGF
Biotecnología
Proliferación Celular
Patología
Protéines de la famille de l'EGF
Biotechnologie
Prolifération cellulaire
Anatomopathologie
Família de Proteínas EGF
Biotecnologia
Proliferação de Células
Patologia
title_short A toolkit for recombinant production of seven human EGF family growth factors in active conformation
title_full A toolkit for recombinant production of seven human EGF family growth factors in active conformation
title_fullStr A toolkit for recombinant production of seven human EGF family growth factors in active conformation
title_full_unstemmed A toolkit for recombinant production of seven human EGF family growth factors in active conformation
title_sort A toolkit for recombinant production of seven human EGF family growth factors in active conformation
author Ferreira, Arthur Schveitzer
author_facet Ferreira, Arthur Schveitzer
Lopacinski, Amanda
Batista, Michel
Hiraiwa, Priscila Mazzocchi
Guimarães, Beatriz Gomes
Zanchin, Nilson Ivo Tonin
author_role author
author2 Lopacinski, Amanda
Batista, Michel
Hiraiwa, Priscila Mazzocchi
Guimarães, Beatriz Gomes
Zanchin, Nilson Ivo Tonin
author2_role author
author
author
author
author
dc.contributor.author.fl_str_mv Ferreira, Arthur Schveitzer
Lopacinski, Amanda
Batista, Michel
Hiraiwa, Priscila Mazzocchi
Guimarães, Beatriz Gomes
Zanchin, Nilson Ivo Tonin
dc.subject.other.pt_BR.fl_str_mv Escherichia coli
topic Escherichia coli
EGF Family of Proteins
Biotechnology
Cell Proliferation
Pathology
Familia de Proteínas EGF
Biotecnología
Proliferación Celular
Patología
Protéines de la famille de l'EGF
Biotechnologie
Prolifération cellulaire
Anatomopathologie
Família de Proteínas EGF
Biotecnologia
Proliferação de Células
Patologia
dc.subject.en.pt_BR.fl_str_mv EGF Family of Proteins
Biotechnology
Cell Proliferation
Pathology
dc.subject.es.pt_BR.fl_str_mv Familia de Proteínas EGF
Biotecnología
Proliferación Celular
Patología
dc.subject.fr.pt_BR.fl_str_mv Protéines de la famille de l'EGF
Biotechnologie
Prolifération cellulaire
Anatomopathologie
dc.subject.decs.pt_BR.fl_str_mv Família de Proteínas EGF
Biotecnologia
Proliferação de Células
Patologia
description Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Biologia Estrutural e Engenharia de Proteínas. Curitiba, PR, Brasil. / Universidade Federal do Paraná. Departamento de Biologia Celular e Molecular. Curitiba, PR, Brasil.
publishDate 2022
dc.date.accessioned.fl_str_mv 2022-05-03T19:42:40Z
dc.date.available.fl_str_mv 2022-05-03T19:42:40Z
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. A toolkit for recombinant production of seven human EGF family growth factors in active conformation. Scientific Reports, v. 12, n. 5034, p. 1–14, 2022.
dc.identifier.uri.fl_str_mv https://www.arca.fiocruz.br/handle/icict/52522
dc.identifier.issn.pt_BR.fl_str_mv 2045-2322
dc.identifier.doi.none.fl_str_mv 10.1038/s41598-022-09060-9
identifier_str_mv FERREIRA, Arthur Schveitzer et al. A toolkit for recombinant production of seven human EGF family growth factors in active conformation. Scientific Reports, v. 12, n. 5034, p. 1–14, 2022.
2045-2322
10.1038/s41598-022-09060-9
url https://www.arca.fiocruz.br/handle/icict/52522
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 Nature
publisher.none.fl_str_mv Nature
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/52522/1/license.txt
https://www.arca.fiocruz.br/bitstream/icict/52522/2/41598_2022_Article_9060OK.pdf
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