Biochemical and Biophysical Characterization of the Caveolin-2 Interaction with Membranes and Analysis of the Protein Structural Alteration by the Presence of Cholesterol

Detalhes bibliográficos
Autor(a) principal: Gorospe, Berta
Data de Publicação: 2022
Outros Autores: Moura, José J. G., Gutierrez-Merino, Carlos, Samhan-Arias, Alejandro K.
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10362/150485
Resumo: This work was supported by the Associate Laboratory for Green Chemistry—LAQV which is financed by national funds from FCT/MCTES (UID/QUI/50006/2020) and co-financed by the ERDF under the PT2020 Partnership Agreement (POCI-01-0145-FEDER—007265). Experimental work was also supported by funding from Ayuda a Grupos de la Junta de Extremadura (Group BBB008-GR21051), co-financed by the European Funds for Structural Development (FEDER). The BioLab is co-financed by the ERDF under the PT2020 Partnership Agreement (POCI-01-0145-FEDER-007728 and POCI-01-0145-FEDER—007265, respectively). Publisher Copyright: © 2022 by the authors.
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spelling Biochemical and Biophysical Characterization of the Caveolin-2 Interaction with Membranes and Analysis of the Protein Structural Alteration by the Presence of Cholesterolcaveolin-1caveolin-2cholesterolmembrane interactionsecondary structure predictionCatalysisMolecular BiologySpectroscopyComputer Science ApplicationsPhysical and Theoretical ChemistryOrganic ChemistryInorganic ChemistryThis work was supported by the Associate Laboratory for Green Chemistry—LAQV which is financed by national funds from FCT/MCTES (UID/QUI/50006/2020) and co-financed by the ERDF under the PT2020 Partnership Agreement (POCI-01-0145-FEDER—007265). Experimental work was also supported by funding from Ayuda a Grupos de la Junta de Extremadura (Group BBB008-GR21051), co-financed by the European Funds for Structural Development (FEDER). The BioLab is co-financed by the ERDF under the PT2020 Partnership Agreement (POCI-01-0145-FEDER-007728 and POCI-01-0145-FEDER—007265, respectively). Publisher Copyright: © 2022 by the authors.Caveolin-2 is a protein suitable for the study of interactions of caveolins with other proteins and lipids present in caveolar lipid rafts. Caveolin-2 has a lower tendency to associate with high molecular weight oligomers than caveolin-1, facilitating the study of its structural modulation upon association with other proteins or lipids. In this paper, we have successfully expressed and purified recombinant human caveolin-2 using E. coli. The structural changes of caveolin-2 upon interaction with a lipid bilayer of liposomes were characterized using bioinformatic prediction models, circular dichroism, differential scanning calorimetry, and fluorescence techniques. Our data support that caveolin-2 binds and alters cholesterol-rich domains in the membranes through a CARC domain, a type of cholesterol-interacting domain in its sequence. The far UV-CD spectra support that the purified protein keeps its folding properties but undergoes a change in its secondary structure in the presence of lipids that correlates with the acquisition of a more stable conformation, as shown by differential scanning calorimetry experiments. Fluorescence experiments using egg yolk lecithin large unilamellar vesicles loaded with 1,6-diphenylhexatriene confirmed that caveolin-2 adsorbs to the membrane but only penetrates the core of the phospholipid bilayer if vesicles are supplemented with 30% of cholesterol. Our study sheds light on the caveolin-2 interaction with lipids. In addition, we propose that purified recombinant caveolin-2 can provide a new tool to study protein–lipid interactions within caveolae.DQ - Departamento de QuímicaLAQV@REQUIMTERUNGorospe, BertaMoura, José J. G.Gutierrez-Merino, CarlosSamhan-Arias, Alejandro K.2023-03-13T22:26:48Z2022-122022-12-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article18application/pdfhttp://hdl.handle.net/10362/150485engGorospe, B., Moura, J. J. G., Gutierrez-Merino, C., & Samhan-Arias, A. K. (2022). Biochemical and Biophysical Characterization of the Caveolin-2 Interaction with Membranes and Analysis of the Protein Structural Alteration by the Presence of Cholesterol. International Journal of Molecular Sciences, 23(23), [15203]. https://doi.org/10.3390/ijms2323152031661-6596PURE: 55548660https://doi.org/10.3390/ijms232315203info: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:RCAAP2024-03-11T05:32:27Zoai:run.unl.pt:10362/150485Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T03:54:08.615260Repositório 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 Biochemical and Biophysical Characterization of the Caveolin-2 Interaction with Membranes and Analysis of the Protein Structural Alteration by the Presence of Cholesterol
title Biochemical and Biophysical Characterization of the Caveolin-2 Interaction with Membranes and Analysis of the Protein Structural Alteration by the Presence of Cholesterol
spellingShingle Biochemical and Biophysical Characterization of the Caveolin-2 Interaction with Membranes and Analysis of the Protein Structural Alteration by the Presence of Cholesterol
Gorospe, Berta
caveolin-1
caveolin-2
cholesterol
membrane interaction
secondary structure prediction
Catalysis
Molecular Biology
Spectroscopy
Computer Science Applications
Physical and Theoretical Chemistry
Organic Chemistry
Inorganic Chemistry
title_short Biochemical and Biophysical Characterization of the Caveolin-2 Interaction with Membranes and Analysis of the Protein Structural Alteration by the Presence of Cholesterol
title_full Biochemical and Biophysical Characterization of the Caveolin-2 Interaction with Membranes and Analysis of the Protein Structural Alteration by the Presence of Cholesterol
title_fullStr Biochemical and Biophysical Characterization of the Caveolin-2 Interaction with Membranes and Analysis of the Protein Structural Alteration by the Presence of Cholesterol
title_full_unstemmed Biochemical and Biophysical Characterization of the Caveolin-2 Interaction with Membranes and Analysis of the Protein Structural Alteration by the Presence of Cholesterol
title_sort Biochemical and Biophysical Characterization of the Caveolin-2 Interaction with Membranes and Analysis of the Protein Structural Alteration by the Presence of Cholesterol
author Gorospe, Berta
author_facet Gorospe, Berta
Moura, José J. G.
Gutierrez-Merino, Carlos
Samhan-Arias, Alejandro K.
author_role author
author2 Moura, José J. G.
Gutierrez-Merino, Carlos
Samhan-Arias, Alejandro K.
author2_role author
author
author
dc.contributor.none.fl_str_mv DQ - Departamento de Química
LAQV@REQUIMTE
RUN
dc.contributor.author.fl_str_mv Gorospe, Berta
Moura, José J. G.
Gutierrez-Merino, Carlos
Samhan-Arias, Alejandro K.
dc.subject.por.fl_str_mv caveolin-1
caveolin-2
cholesterol
membrane interaction
secondary structure prediction
Catalysis
Molecular Biology
Spectroscopy
Computer Science Applications
Physical and Theoretical Chemistry
Organic Chemistry
Inorganic Chemistry
topic caveolin-1
caveolin-2
cholesterol
membrane interaction
secondary structure prediction
Catalysis
Molecular Biology
Spectroscopy
Computer Science Applications
Physical and Theoretical Chemistry
Organic Chemistry
Inorganic Chemistry
description This work was supported by the Associate Laboratory for Green Chemistry—LAQV which is financed by national funds from FCT/MCTES (UID/QUI/50006/2020) and co-financed by the ERDF under the PT2020 Partnership Agreement (POCI-01-0145-FEDER—007265). Experimental work was also supported by funding from Ayuda a Grupos de la Junta de Extremadura (Group BBB008-GR21051), co-financed by the European Funds for Structural Development (FEDER). The BioLab is co-financed by the ERDF under the PT2020 Partnership Agreement (POCI-01-0145-FEDER-007728 and POCI-01-0145-FEDER—007265, respectively). Publisher Copyright: © 2022 by the authors.
publishDate 2022
dc.date.none.fl_str_mv 2022-12
2022-12-01T00:00:00Z
2023-03-13T22:26:48Z
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/10362/150485
url http://hdl.handle.net/10362/150485
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Gorospe, B., Moura, J. J. G., Gutierrez-Merino, C., & Samhan-Arias, A. K. (2022). Biochemical and Biophysical Characterization of the Caveolin-2 Interaction with Membranes and Analysis of the Protein Structural Alteration by the Presence of Cholesterol. International Journal of Molecular Sciences, 23(23), [15203]. https://doi.org/10.3390/ijms232315203
1661-6596
PURE: 55548660
https://doi.org/10.3390/ijms232315203
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
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application/pdf
dc.source.none.fl_str_mv reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
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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
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