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Structural analysis of the GPI glycan

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dc.contributor.authorNakano, Miyako
dc.contributor.authorSabido-Bozo, Susana
dc.contributor.authorOkazaki, Kouta
dc.contributor.authorAguilera-Romero, Auxiliadora
dc.contributor.authorRodriguez-Gallardo, Sofia
dc.contributor.authorCortes-Gomez, Alejandro
dc.contributor.authorLopez, Sergio
dc.contributor.authorIkeda, Atsuko
dc.contributor.authorFunato, Kouichi
dc.contributor.authorMuñiz, Manuel
dc.contributor.authoraffiliation[Nakano,M; Okazaki,K; Ikeda,A; Funato,K] Graduate School of Integrated Sciences for Life, Hiroshima University, Higashi-Hiroshima, Hiroshima, Japan. [Sabido-Bozo,S; Aguilera-Romero,A; Rodriguez-Gallardo,S; Cortes-Gomez,A; Lopez,S; Muñiz,M] Facultad de Biologia, Departamento de Biologia Celular, Hospital Universitario Virgen del Rocio/CSIC/Universidad de Sevilla, Universidad de Sevilla e Instituto de Biomedicina de Sevilla (IBiS), Sevilla, Spain.
dc.contributor.funderMM. BFU2017-89700-P. FEDER/Ministerio de Ciencia, Innovación y Universidades—Agencia Estatal de Investigación. http://www.aei.gob.es/. MN and KF. JP19H02922. Japan Society for the Promotion of Science. https://www.jsps.go.jp/english/. The funders had and will not have a role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
dc.date.accessioned2022-11-25T16:09:13Z
dc.date.available2022-11-25T16:09:13Z
dc.date.issued2021-09-16
dc.description.abstractGlycosylphosphatidylinositol (GPI) anchoring of proteins is an essential post-translational modification in all eukaryotes that occurs at the endoplasmic reticulum (ER) and serves to deliver GPI-anchored proteins (GPI-APs) to the cell surface where they play a wide variety of vital physiological roles. This paper describes a specialized method for purification and structural analysis of the GPI glycan of individual GPI-APs in yeast. The protocol involves the expression of a specific GPI-AP tagged with GFP, enzymatic release from the cellular membrane fraction, immunopurification, separation by electrophoresis and analysis of the peptides bearing GPI glycans by mass spectrometry after trypsin digestion. We used specifically this protocol to address the structural remodeling that undergoes the GPI glycan of a specific GPI-AP during its transport to the cell surface. This method can be also applied to investigate the GPI-AP biosynthetic pathway and to directly confirm predicted GPI-anchoring of individual proteins.es_ES
dc.description.versionYeses_ES
dc.identifier.citationNakano M, Sabido-Bozo S, Okazaki K, Aguilera-Romero A, Rodriguez-Gallardo S, Cortes-Gomez A, et al. Structural analysis of the GPI glycan. PLoS One. 2021 Sep 16;16(9):e0257435es_ES
dc.identifier.doi10.1371/journal.pone.0257435es_ES
dc.identifier.essn1932-6203
dc.identifier.pmcPMC8445438
dc.identifier.pmid34529709es_ES
dc.identifier.urihttp://hdl.handle.net/10668/4410
dc.journal.titlePLoS One
dc.language.isoen
dc.page.number6 p.
dc.publisherPublic Library of Sciencees_ES
dc.relation.publisherversionhttps://journals.plos.org/plosone/article?id=10.1371/journal.pone.0257435es_ES
dc.rightsAtribución 4.0 Internacional*
dc.rights.accessRightsopen access
dc.rights.urihttp://creativecommons.org/licenses/by/4.0/*
dc.subjectGlycosylphosphatidylinositoles_ES
dc.subjectTrypsines_ES
dc.subjectEndoplasmic reticulumes_ES
dc.subjectMass spectrometryes_ES
dc.subjectElectrophoresises_ES
dc.subjectDigestiones_ES
dc.subjectPeptideses_ES
dc.subjectGlicosilfosfatidilinositoleses_ES
dc.subjectTripsinaes_ES
dc.subjectRetículo endoplásmicoes_ES
dc.subjectEspectrometría de masases_ES
dc.subjectElectroforesises_ES
dc.subjectDigestiónes_ES
dc.subjectPéptidoses_ES
dc.subject.meshMedical Subject Headings::Anatomy::Cells::Cellular Structures::Intracellular Space::Cytoplasm::Cytoplasmic Structures::Organelles::Endoplasmic Reticulumes_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::Chemicals and Drugs::Amino Acids, Peptides, and Proteins::Peptideses_ES
dc.subject.meshMedical Subject Headings::Organisms::Eukaryota::Fungi::Ascomycota::Saccharomycetales::Saccharomyces::Saccharomyces cerevisiaees_ES
dc.subject.meshMedical Subject Headings::Analytical, Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Chemistry Techniques, Analytical::Mass Spectrometry::Tandem Mass Spectrometryes_ES
dc.subject.meshMedical Subject Headings::Chemicals and Drugs::Carbohydrates::Glycoconjugates::Glycolipids::Glycosylphosphatidylinositolses_ES
dc.subject.meshMedical Subject Headings::Chemicals and Drugs::Enzymes and Coenzymes::Enzymes::Hydrolases::Peptide Hydrolases::Endopeptidases::Serine Endopeptidases::Trypsines_ES
dc.subject.meshMedical Subject Headings::Phenomena and Processes::Chemical Phenomena::Biochemical Phenomena::Biochemical Processes::Biosynthetic Pathwayses_ES
dc.subject.meshMedical Subject Headings::Phenomena and Processes::Chemical Phenomena::Biochemical Phenomena::Biochemical Processes::Protein Modification, Translational::Protein Processing, Post-Translationales_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::Analytical, Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Chemistry Techniques, Analytical::Electrophoresises_ES
dc.subject.meshMedical Subject Headings::Phenomena and Processes::Digestive System and Oral Physiological Phenomena::Digestive System Physiological Phenomena::Digestive System Processes::Digestiones_ES
dc.titleStructural analysis of the GPI glycanes_ES
dc.typeresearch article
dc.type.hasVersionVoR
dspace.entity.typePublication

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