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Determination of the lipid composition of the GPI anchor

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dc.contributor.authorAguilera-Romero, Auxiliadora
dc.contributor.authorSabido-Bozo, Susana
dc.contributor.authorLopez, Sergio
dc.contributor.authorCortes-Gomez, Alejandro
dc.contributor.authorRodriguez-Gallardo, Sofia
dc.contributor.authorPerez-Linero, Ana Maria
dc.contributor.authorRiezman, Isabelle
dc.contributor.authorRiezman, Howard
dc.contributor.authorMuñiz, Manuel
dc.contributor.authoraffiliation[Aguilera-Romero,A; Sabido-Bozo,S; Lopez,S; Cortes-Gomez,A; Rodriguez-Gallardo,S; Perez-Linero,AM; Muñiz,M] Departamento de Biología Celular, Facultad de Biología, Universidad de Sevilla e Instituto de Biomedicina de Sevilla (IBiS), Hospital Universitario Virgen del Rocío/CSIC/Universidad de Sevilla, Seville, Spain. [Riezman,I; Riezman,H] Department of Biochemistry, NCCR Chemical Biology, University of Geneva, Geneva, Switzerland.
dc.contributor.funderThis research was funded by the FEDER/Ministerio de Ciencia, Innovación y Universidades—Agencia Estatal de Investigación/BFU2017-89700-P to Manuel Muñiz, "VI Own Research Plan" of the University of Seville VIPPIT-2020-I.5 to Manuel Muñiz, by the Incentivo al Grupo de Investigación BIO-271 (2019/BIO-271) and the NCCR Chemical Biology and the Swiss National Science Foundation (51NF40-185898 and 310030_184949) to Howard Riezman. ‘V Own Research Plan’ of the University of Seville (VPPIUS) contract (cofounded by the European Social Fund) to Sergio Lopez, University of Seville fellowships to Sofia Rodriguez-Gallardo and Ana Maria Perez-Linero, Ministry of Education, Culture and Sport (MECD) fellowship to Susana Sabido Bozo and contract from the University of Seville by the Youth Employment Initiative to Alejandro Cortes-Gomez.
dc.date.accessioned2022-10-31T13:31:07Z
dc.date.available2022-10-31T13:31:07Z
dc.date.issued2021-08-13
dc.description.abstractIn eukaryotic cells, a subset of cell surface proteins is attached by the glycolipid glycosylphosphatidylinositol (GPI) to the external leaflet of the plasma membrane where they play important roles as enzymes, receptors, or adhesion molecules. Here we present a protocol for purification and mass spectrometry analysis of the lipid moiety of individual GPI-anchored proteins (GPI-APs) in yeast. The method involves the expression of a specific GPI-AP tagged with GFP, solubilization, immunoprecipitation, separation by electrophoresis, blotting onto PVDF, release and extraction of the GPI-lipid moiety and analysis by mass spectrometry. By using this protocol, we could determine the precise GPI-lipid structure of the GPI-AP Gas1-GFP in a modified yeast strain. This protocol can be used to identify the lipid composition of the GPI anchor of distinct GPI-APs from yeast to mammals and can be adapted to determine other types of protein lipidation.es_ES
dc.description.versionYeses_ES
dc.identifier.citationAguilera-Romero A, Sabido-Bozo S, Lopez S, Cortes-Gomez A, Rodriguez-Gallardo S, Perez-Linero AM, et al. Determination of the lipid composition of the GPI anchor. PLoS One. 2021 Aug 13;16(8):e0256184es_ES
dc.identifier.doi10.1371/journal.pone.0256184es_ES
dc.identifier.essn1932-6203
dc.identifier.pmcPMC8362999
dc.identifier.pmid34388214es_ES
dc.identifier.urihttp://hdl.handle.net/10668/4314
dc.journal.titlePloS One
dc.language.isoen
dc.page.number5 p.
dc.publisherPublic Library of Sciencees_ES
dc.relation.publisherversionhttps://journals.plos.org/plosone/article?id=10.1371/journal.pone.0256184es_ES
dc.rightsAtribución 4.0 Internacional*
dc.rights.accessRightsopen access
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/*
dc.subjectLipidses_ES
dc.subjectGlycosylphosphatidylinositolses_ES
dc.subjectMembraneses_ES
dc.subjectCeramideses_ES
dc.subjectGenes, reporteres_ES
dc.subjectLípidoses_ES
dc.subjectGlicosilfosfatidilinositoleses_ES
dc.subjectMembranases_ES
dc.subjectCeramidases_ES
dc.subjectGenes informadoreses_ES
dc.subject.meshMedical Subject Headings::Anatomy::Cells::Cellular Structures::Cell Membranees_ES
dc.subject.meshMedical Subject Headings::Chemicals and Drugs::Organic Chemicals::Amides::Ceramideses_ES
dc.subject.meshMedical Subject Headings::Chemicals and Drugs::Amino Acids, Peptides, and Proteins::Proteins::Glycoproteins::Membrane Glycoproteins::GPI-Linked Proteinses_ES
dc.subject.meshMedical Subject Headings::Phenomena and Processes::Genetic Phenomena::Genetic Structures::Genome::Genome Components::Genes::Genes, Reporteres_ES
dc.subject.meshMedical Subject Headings::Chemicals and Drugs::Carbohydrates::Glycoconjugates::Glycolipids::Glycosylphosphatidylinositolses_ES
dc.subject.meshMedical Subject Headings::Chemicals and Drugs::Amino Acids, Peptides, and Proteins::Proteins::Luminescent Proteins::Green Fluorescent Proteinses_ES
dc.subject.meshMedical Subject Headings::Analytical, Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Chemistry Techniques, Analytical::Mass Spectrometryes_ES
dc.subject.meshMedical Subject Headings::Chemicals and Drugs::Amino Acids, Peptides, and Proteins::Proteins::Glycoproteins::Membrane Glycoproteinses_ES
dc.subject.meshMedical Subject Headings::Organisms::Eukaryota::Fungi::Ascomycota::Saccharomycetales::Saccharomyceses_ES
dc.subject.meshMedical Subject Headings::Chemicals and Drugs::Amino Acids, Peptides, and Proteins::Proteins::Fungal Proteins::Saccharomyces cerevisiae Proteinses_ES
dc.titleDetermination of the lipid composition of the GPI anchores_ES
dc.typeresearch article
dc.type.hasVersionVoR
dspace.entity.typePublication

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