Enhanced stability of detergent-free human native STEAP1 protein from neoplastic prostate cancer cells upon an innovative isolation procedure

Detalhes bibliográficos
Autor(a) principal: Barroca-Ferreira, Jorge
Data de Publicação: 2021
Outros Autores: Cruz-Vicente, Pedro, Santos, Marino F. A., Rocha, Sandra M., Santos-Silva, Teresa, Maia, Cláudio J., Passarinha, Luís A.
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10362/142769
Resumo: POCI‐01‐0145‐FEDER‐ 007491 LA/P/0140/2020
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spelling Enhanced stability of detergent-free human native STEAP1 protein from neoplastic prostate cancer cells upon an innovative isolation procedureCircular dichroismCo-immunoprecipitationProstate cancerProtein purificationSTEAP1Thermal stabilityCatalysisMolecular BiologySpectroscopyComputer Science ApplicationsPhysical and Theoretical ChemistryOrganic ChemistryInorganic ChemistrySDG 3 - Good Health and Well-beingPOCI‐01‐0145‐FEDER‐ 007491 LA/P/0140/2020Background: The STEAP1 is a cell-surface antigen over-expressed in prostate cancer, which contributes to tumor progression and aggressiveness. However, the molecular mechanisms underlying STEAP1 and its structural determinants remain elusive. Methods: The fraction capacity of Butyl- and Octyl-Sepharose matrices on LNCaP lysates was evaluated by manipulating the ionic strength of binding and elution phases, followed by a Co-Immunoprecipitation (Co-IP) polishing. Several potential stabilizing additives were assessed, and the melting temperature (Tm) values ranked the best/worst compounds. The secondary structure of STEAP1 was identified by circular dichroism. Results: The STEAP1 was not fully captured with 1.375 M (Butyl), in contrast with interfering heterologous proteins, which were strongly retained and mostly eluted with water. This single step demonstrated higher selectivity of Butyl-Sepharose for host impurities removal from injected crude samples. Co-IP allowed recovering a purified fraction of STEAP1 and contributed to unveil potential physiologically interacting counterparts with the target. A Tm of ~55 °C was determined, confirming STEAP1 stability in the purification buffer. A predominant α-helical structure was identified, ensuring the protein’s structural stability. Conclusions: A method for successfully isolating human STEAP1 from LNCaP cells was provided, avoiding the use of detergents to achieve stability, even outside a membrane-mimicking environment.UCIBIO - Applied Molecular Biosciences UnitDQ - Departamento de QuímicaRUNBarroca-Ferreira, JorgeCruz-Vicente, PedroSantos, Marino F. A.Rocha, Sandra M.Santos-Silva, TeresaMaia, Cláudio J.Passarinha, Luís A.2022-08-01T22:25:51Z2021-092021-09-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10362/142769eng1661-6596PURE: 45749340https://doi.org/10.3390/ijms221810012info: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:RCAAP2024-03-11T05:20:40Zoai:run.unl.pt:10362/142769Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T03:50:30.565279Repositório 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 Enhanced stability of detergent-free human native STEAP1 protein from neoplastic prostate cancer cells upon an innovative isolation procedure
title Enhanced stability of detergent-free human native STEAP1 protein from neoplastic prostate cancer cells upon an innovative isolation procedure
spellingShingle Enhanced stability of detergent-free human native STEAP1 protein from neoplastic prostate cancer cells upon an innovative isolation procedure
Barroca-Ferreira, Jorge
Circular dichroism
Co-immunoprecipitation
Prostate cancer
Protein purification
STEAP1
Thermal stability
Catalysis
Molecular Biology
Spectroscopy
Computer Science Applications
Physical and Theoretical Chemistry
Organic Chemistry
Inorganic Chemistry
SDG 3 - Good Health and Well-being
title_short Enhanced stability of detergent-free human native STEAP1 protein from neoplastic prostate cancer cells upon an innovative isolation procedure
title_full Enhanced stability of detergent-free human native STEAP1 protein from neoplastic prostate cancer cells upon an innovative isolation procedure
title_fullStr Enhanced stability of detergent-free human native STEAP1 protein from neoplastic prostate cancer cells upon an innovative isolation procedure
title_full_unstemmed Enhanced stability of detergent-free human native STEAP1 protein from neoplastic prostate cancer cells upon an innovative isolation procedure
title_sort Enhanced stability of detergent-free human native STEAP1 protein from neoplastic prostate cancer cells upon an innovative isolation procedure
author Barroca-Ferreira, Jorge
author_facet Barroca-Ferreira, Jorge
Cruz-Vicente, Pedro
Santos, Marino F. A.
Rocha, Sandra M.
Santos-Silva, Teresa
Maia, Cláudio J.
Passarinha, Luís A.
author_role author
author2 Cruz-Vicente, Pedro
Santos, Marino F. A.
Rocha, Sandra M.
Santos-Silva, Teresa
Maia, Cláudio J.
Passarinha, Luís A.
author2_role author
author
author
author
author
author
dc.contributor.none.fl_str_mv UCIBIO - Applied Molecular Biosciences Unit
DQ - Departamento de Química
RUN
dc.contributor.author.fl_str_mv Barroca-Ferreira, Jorge
Cruz-Vicente, Pedro
Santos, Marino F. A.
Rocha, Sandra M.
Santos-Silva, Teresa
Maia, Cláudio J.
Passarinha, Luís A.
dc.subject.por.fl_str_mv Circular dichroism
Co-immunoprecipitation
Prostate cancer
Protein purification
STEAP1
Thermal stability
Catalysis
Molecular Biology
Spectroscopy
Computer Science Applications
Physical and Theoretical Chemistry
Organic Chemistry
Inorganic Chemistry
SDG 3 - Good Health and Well-being
topic Circular dichroism
Co-immunoprecipitation
Prostate cancer
Protein purification
STEAP1
Thermal stability
Catalysis
Molecular Biology
Spectroscopy
Computer Science Applications
Physical and Theoretical Chemistry
Organic Chemistry
Inorganic Chemistry
SDG 3 - Good Health and Well-being
description POCI‐01‐0145‐FEDER‐ 007491 LA/P/0140/2020
publishDate 2021
dc.date.none.fl_str_mv 2021-09
2021-09-01T00:00:00Z
2022-08-01T22:25:51Z
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/10362/142769
url http://hdl.handle.net/10362/142769
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 1661-6596
PURE: 45749340
https://doi.org/10.3390/ijms221810012
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.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
instname_str 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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