Author | Ferreira, Arthur Schveitzer | |
Author | Lopacinski, Amanda | |
Author | Batista, Michel | |
Author | Hiraiwa, Priscila Mazzocchi | |
Author | Guimarães, Beatriz Gomes | |
Author | Zanchin, Nilson Ivo Tonin | |
Access date | 2022-05-03T19:42:40Z | |
Available date | 2022-05-03T19:42:40Z | |
Document date | 2022 | |
Citation | 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. | pt_BR |
ISSN | 2045-2322 | pt_BR |
URI | https://www.arca.fiocruz.br/handle/icict/52522 | |
Language | por | pt_BR |
Publisher | Nature | pt_BR |
Rights | open access | pt_BR |
Subject in Portuguese | Escherichia coli | pt_BR |
Title | A toolkit for recombinant production of seven human EGF family growth factors in active conformation | pt_BR |
Type | Article | pt_BR |
DOI | 10.1038/s41598-022-09060-9 | |
Abstract | 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. | pt_BR |
Affilliation | 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. | pt_BR |
Affilliation | 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. | pt_BR |
Affilliation | Fundação Oswaldo Cruz. Instituto Carlos Chagas. Mass Spectrometry Facility - RPT02H. Curitiba, PR, Brasil. | pt_BR |
Affilliation | Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Biologia Estrutural e Engenharia de Proteínas. Curitiba, PR, Brasil. | pt_BR |
Affilliation | Fundação Oswaldo Cruz. Instituto Carlos Chagas. Laboratório de Biologia Estrutural e Engenharia de Proteínas. Curitiba, PR, Brasil. | pt_BR |
Affilliation | 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. | pt_BR |
Subject | EGF Family of Proteins | pt_BR |
Subject | Biotechnology | pt_BR |
Subject | Cell Proliferation | pt_BR |
Subject | Pathology | pt_BR |
Subject in Spanish | Familia de Proteínas EGF | pt_BR |
Subject in Spanish | Biotecnología | pt_BR |
Subject in Spanish | Proliferación Celular | pt_BR |
Subject in Spanish | Patología | pt_BR |
Subject in French | Protéines de la famille de l'EGF | pt_BR |
Subject in French | Biotechnologie | pt_BR |
Subject in French | Prolifération cellulaire | pt_BR |
Subject in French | Anatomopathologie | pt_BR |
DeCS | Família de Proteínas EGF | pt_BR |
DeCS | Biotecnologia | pt_BR |
DeCS | Proliferação de Células | pt_BR |
DeCS | Patologia | pt_BR |