The Diacylglycerol Acyltransferase 3 of Chlamydomonas reinhardtii Is a Disordered Protein Capable of Binding to Lipids Derived from Chloroplasts

Autores
Pavía, Natalia; Potenza, Alberto; Hornos, Felipe; Poveda, José A.; Gonorazky, Ana Gabriela; Neira, José L.; Giudici, Ana Marcela; Beligni, María Verónica
Año de publicación
2025
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Understanding triacylglycerol (TAG) metabolism is crucial for developing algae as a source of biodiesel. TAGs are the main reservoir of energy in most eukaryotes. The final, rate-limiting step in the formation of TAGs is catalyzed by 1,2-diacylglycerol acyltransferases (DGATs). In the green alga Chlamydomonas reinhardtii, DGAT3 is phylogenetically related to plant DGAT3 but unrelated to other DGATs from eukaryotes, such as DGAT1 and DGAT2. In this study, we described the conformational preferences and the lipid-binding features of the DGAT3 from C. reinhardtii. To characterize its conformational stability and structural features, we used several biophysical probes, namely, fluorescence, circular dichroism (CD), and differential scanning calorimetry (DSC). Our results showed that the protein was mainly disordered, containing a small population of folded conformations in a narrow pH range (pH 8 to 10). The conformational stability of the folded structure of DGAT3 was very low, as shown by urea or guanidinium denaturations. Thermal denaturation, followed by fluorescence or CD, as well as calorimetric denaturation, followed by DSC, did not yield any transition in the pH range where DGAT3 acquired a “native-like” conformation. Furthermore, we used two approaches to demonstrate the interaction of DGAT3 with lipid membranes at the pH at which it had acquired a “native-like” conformation. The first involved the measurement of anisotropy and fluorescence quenching of the protein. The second approach focused on examining possible modifications of the biophysical properties of lipids due to their interaction with DGAT3, through anisotropy measurements and leakage assays. Both methods produced consistent results, suggesting that DGAT3 preferentially interacted with negatively charged membranes. These results will allow the design of a more efficient and stable DGAT3, as well as an in-depth understanding of how the metabolism of TAGs is accomplished in C. reinhardtii.
Fil: Pavía, Natalia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentina
Fil: Potenza, Alberto. Universidad de Miguel Hernández; España
Fil: Hornos, Felipe. Universidad de Miguel Hernández; España
Fil: Poveda, José A.. Universidad de Miguel Hernández; España
Fil: Gonorazky, Ana Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentina
Fil: Neira, José L.. Universidad de Miguel Hernández; España
Fil: Giudici, Ana Marcela. Universidad de Miguel Hernández; España. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Beligni, María Verónica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentina
Materia
PROTEIN-LIPID INTERACTION
LEAKAGE
FLUORESCENCE
DISORDERED PROTEIN
THYLAKOIDS
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/289710
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/289710
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repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling The Diacylglycerol Acyltransferase 3 of Chlamydomonas reinhardtii Is a Disordered Protein Capable of Binding to Lipids Derived from ChloroplastsPavía, NataliaPotenza, AlbertoHornos, FelipePoveda, José A.Gonorazky, Ana GabrielaNeira, José L.Giudici, Ana MarcelaBeligni, María VerónicaPROTEIN-LIPID INTERACTIONLEAKAGEFLUORESCENCEDISORDERED PROTEINTHYLAKOIDShttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Understanding triacylglycerol (TAG) metabolism is crucial for developing algae as a source of biodiesel. TAGs are the main reservoir of energy in most eukaryotes. The final, rate-limiting step in the formation of TAGs is catalyzed by 1,2-diacylglycerol acyltransferases (DGATs). In the green alga Chlamydomonas reinhardtii, DGAT3 is phylogenetically related to plant DGAT3 but unrelated to other DGATs from eukaryotes, such as DGAT1 and DGAT2. In this study, we described the conformational preferences and the lipid-binding features of the DGAT3 from C. reinhardtii. To characterize its conformational stability and structural features, we used several biophysical probes, namely, fluorescence, circular dichroism (CD), and differential scanning calorimetry (DSC). Our results showed that the protein was mainly disordered, containing a small population of folded conformations in a narrow pH range (pH 8 to 10). The conformational stability of the folded structure of DGAT3 was very low, as shown by urea or guanidinium denaturations. Thermal denaturation, followed by fluorescence or CD, as well as calorimetric denaturation, followed by DSC, did not yield any transition in the pH range where DGAT3 acquired a “native-like” conformation. Furthermore, we used two approaches to demonstrate the interaction of DGAT3 with lipid membranes at the pH at which it had acquired a “native-like” conformation. The first involved the measurement of anisotropy and fluorescence quenching of the protein. The second approach focused on examining possible modifications of the biophysical properties of lipids due to their interaction with DGAT3, through anisotropy measurements and leakage assays. Both methods produced consistent results, suggesting that DGAT3 preferentially interacted with negatively charged membranes. These results will allow the design of a more efficient and stable DGAT3, as well as an in-depth understanding of how the metabolism of TAGs is accomplished in C. reinhardtii.Fil: Pavía, Natalia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; ArgentinaFil: Potenza, Alberto. Universidad de Miguel Hernández; EspañaFil: Hornos, Felipe. Universidad de Miguel Hernández; EspañaFil: Poveda, José A.. Universidad de Miguel Hernández; EspañaFil: Gonorazky, Ana Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; ArgentinaFil: Neira, José L.. Universidad de Miguel Hernández; EspañaFil: Giudici, Ana Marcela. Universidad de Miguel Hernández; España. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Beligni, María Verónica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; ArgentinaMDPI2025-02info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/289710Pavía, Natalia; Potenza, Alberto; Hornos, Felipe; Poveda, José A.; Gonorazky, Ana Gabriela; et al.; The Diacylglycerol Acyltransferase 3 of Chlamydomonas reinhardtii Is a Disordered Protein Capable of Binding to Lipids Derived from Chloroplasts; MDPI; Biomolecules; 15; 2; 2-2025; 1-232218-273XCONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.mdpi.com/2218-273X/15/2/245info:eu-repo/semantics/altIdentifier/doi/10.3390/biom15020245info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2026-08-25T15:12:23Zoai:ri.conicet.gov.ar:11336/289710instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982026-08-25 15:12:23.658CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv The Diacylglycerol Acyltransferase 3 of Chlamydomonas reinhardtii Is a Disordered Protein Capable of Binding to Lipids Derived from Chloroplasts
title The Diacylglycerol Acyltransferase 3 of Chlamydomonas reinhardtii Is a Disordered Protein Capable of Binding to Lipids Derived from Chloroplasts
spellingShingle The Diacylglycerol Acyltransferase 3 of Chlamydomonas reinhardtii Is a Disordered Protein Capable of Binding to Lipids Derived from Chloroplasts
Pavía, Natalia
PROTEIN-LIPID INTERACTION
LEAKAGE
FLUORESCENCE
DISORDERED PROTEIN
THYLAKOIDS
title_short The Diacylglycerol Acyltransferase 3 of Chlamydomonas reinhardtii Is a Disordered Protein Capable of Binding to Lipids Derived from Chloroplasts
title_full The Diacylglycerol Acyltransferase 3 of Chlamydomonas reinhardtii Is a Disordered Protein Capable of Binding to Lipids Derived from Chloroplasts
title_fullStr The Diacylglycerol Acyltransferase 3 of Chlamydomonas reinhardtii Is a Disordered Protein Capable of Binding to Lipids Derived from Chloroplasts
title_full_unstemmed The Diacylglycerol Acyltransferase 3 of Chlamydomonas reinhardtii Is a Disordered Protein Capable of Binding to Lipids Derived from Chloroplasts
title_sort The Diacylglycerol Acyltransferase 3 of Chlamydomonas reinhardtii Is a Disordered Protein Capable of Binding to Lipids Derived from Chloroplasts
dc.creator.none.fl_str_mv Pavía, Natalia
Potenza, Alberto
Hornos, Felipe
Poveda, José A.
Gonorazky, Ana Gabriela
Neira, José L.
Giudici, Ana Marcela
Beligni, María Verónica
author Pavía, Natalia
author_facet Pavía, Natalia
Potenza, Alberto
Hornos, Felipe
Poveda, José A.
Gonorazky, Ana Gabriela
Neira, José L.
Giudici, Ana Marcela
Beligni, María Verónica
author_role author
author2 Potenza, Alberto
Hornos, Felipe
Poveda, José A.
Gonorazky, Ana Gabriela
Neira, José L.
Giudici, Ana Marcela
Beligni, María Verónica
author2_role author
author
author
author
author
author
author
dc.subject.none.fl_str_mv PROTEIN-LIPID INTERACTION
LEAKAGE
FLUORESCENCE
DISORDERED PROTEIN
THYLAKOIDS
topic PROTEIN-LIPID INTERACTION
LEAKAGE
FLUORESCENCE
DISORDERED PROTEIN
THYLAKOIDS
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Understanding triacylglycerol (TAG) metabolism is crucial for developing algae as a source of biodiesel. TAGs are the main reservoir of energy in most eukaryotes. The final, rate-limiting step in the formation of TAGs is catalyzed by 1,2-diacylglycerol acyltransferases (DGATs). In the green alga Chlamydomonas reinhardtii, DGAT3 is phylogenetically related to plant DGAT3 but unrelated to other DGATs from eukaryotes, such as DGAT1 and DGAT2. In this study, we described the conformational preferences and the lipid-binding features of the DGAT3 from C. reinhardtii. To characterize its conformational stability and structural features, we used several biophysical probes, namely, fluorescence, circular dichroism (CD), and differential scanning calorimetry (DSC). Our results showed that the protein was mainly disordered, containing a small population of folded conformations in a narrow pH range (pH 8 to 10). The conformational stability of the folded structure of DGAT3 was very low, as shown by urea or guanidinium denaturations. Thermal denaturation, followed by fluorescence or CD, as well as calorimetric denaturation, followed by DSC, did not yield any transition in the pH range where DGAT3 acquired a “native-like” conformation. Furthermore, we used two approaches to demonstrate the interaction of DGAT3 with lipid membranes at the pH at which it had acquired a “native-like” conformation. The first involved the measurement of anisotropy and fluorescence quenching of the protein. The second approach focused on examining possible modifications of the biophysical properties of lipids due to their interaction with DGAT3, through anisotropy measurements and leakage assays. Both methods produced consistent results, suggesting that DGAT3 preferentially interacted with negatively charged membranes. These results will allow the design of a more efficient and stable DGAT3, as well as an in-depth understanding of how the metabolism of TAGs is accomplished in C. reinhardtii.
Fil: Pavía, Natalia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentina
Fil: Potenza, Alberto. Universidad de Miguel Hernández; España
Fil: Hornos, Felipe. Universidad de Miguel Hernández; España
Fil: Poveda, José A.. Universidad de Miguel Hernández; España
Fil: Gonorazky, Ana Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentina
Fil: Neira, José L.. Universidad de Miguel Hernández; España
Fil: Giudici, Ana Marcela. Universidad de Miguel Hernández; España. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Beligni, María Verónica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; Argentina
description Understanding triacylglycerol (TAG) metabolism is crucial for developing algae as a source of biodiesel. TAGs are the main reservoir of energy in most eukaryotes. The final, rate-limiting step in the formation of TAGs is catalyzed by 1,2-diacylglycerol acyltransferases (DGATs). In the green alga Chlamydomonas reinhardtii, DGAT3 is phylogenetically related to plant DGAT3 but unrelated to other DGATs from eukaryotes, such as DGAT1 and DGAT2. In this study, we described the conformational preferences and the lipid-binding features of the DGAT3 from C. reinhardtii. To characterize its conformational stability and structural features, we used several biophysical probes, namely, fluorescence, circular dichroism (CD), and differential scanning calorimetry (DSC). Our results showed that the protein was mainly disordered, containing a small population of folded conformations in a narrow pH range (pH 8 to 10). The conformational stability of the folded structure of DGAT3 was very low, as shown by urea or guanidinium denaturations. Thermal denaturation, followed by fluorescence or CD, as well as calorimetric denaturation, followed by DSC, did not yield any transition in the pH range where DGAT3 acquired a “native-like” conformation. Furthermore, we used two approaches to demonstrate the interaction of DGAT3 with lipid membranes at the pH at which it had acquired a “native-like” conformation. The first involved the measurement of anisotropy and fluorescence quenching of the protein. The second approach focused on examining possible modifications of the biophysical properties of lipids due to their interaction with DGAT3, through anisotropy measurements and leakage assays. Both methods produced consistent results, suggesting that DGAT3 preferentially interacted with negatively charged membranes. These results will allow the design of a more efficient and stable DGAT3, as well as an in-depth understanding of how the metabolism of TAGs is accomplished in C. reinhardtii.
publishDate 2025
dc.date.none.fl_str_mv 2025-02
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/289710
Pavía, Natalia; Potenza, Alberto; Hornos, Felipe; Poveda, José A.; Gonorazky, Ana Gabriela; et al.; The Diacylglycerol Acyltransferase 3 of Chlamydomonas reinhardtii Is a Disordered Protein Capable of Binding to Lipids Derived from Chloroplasts; MDPI; Biomolecules; 15; 2; 2-2025; 1-23
2218-273X
CONICET Digital
CONICET
url http://hdl.handle.net/11336/289710
identifier_str_mv Pavía, Natalia; Potenza, Alberto; Hornos, Felipe; Poveda, José A.; Gonorazky, Ana Gabriela; et al.; The Diacylglycerol Acyltransferase 3 of Chlamydomonas reinhardtii Is a Disordered Protein Capable of Binding to Lipids Derived from Chloroplasts; MDPI; Biomolecules; 15; 2; 2-2025; 1-23
2218-273X
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://www.mdpi.com/2218-273X/15/2/245
info:eu-repo/semantics/altIdentifier/doi/10.3390/biom15020245
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv MDPI
publisher.none.fl_str_mv MDPI
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 13.265058

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