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Structural analysis of the GPI glycan
by
Funato, Kouichi
, Cortés Gómez, Alejandro
, Universidad de Sevilla. Departamento de Biología Celular
, Rodríguez Gallardo, Sofía
, Okazaki, Kouta
, European Commission (EC). Fondo Europeo de Desarrollo Regional (FEDER)
, Muñiz Guinea, Manuel
, López Martín, Sergio
, Aguilera Romero, María Auxiliadora
, Ministerio de Ciencia, Innovación y Universidades (MICINN). España
, Ikeda, Atsuko
, Nakano, Miyako
, Japan Society for the Promotion of Science
, Sabido Bozo, Susana
in
Analysis
/ Biology and Life Sciences
/ Cell membranes
/ Cell surface
/ Electrophoresis
/ Endoplasmic reticulum
/ Enzymes
/ Eukaryotes
/ Glycan
/ Glycolipids
/ Glycosylphosphatidylinositol
/ Ions
/ Lab Protocol
/ Mass spectrometry
/ Mass spectroscopy
/ Peptides
/ Plasma
/ Polysaccharides
/ Post-translation
/ Proteins
/ Research and Analysis Methods
/ Scientific imaging
/ Structural analysis
/ Structure
/ Trypsin
/ Yeast
/ Yeasts
2021
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Structural analysis of the GPI glycan
by
Funato, Kouichi
, Cortés Gómez, Alejandro
, Universidad de Sevilla. Departamento de Biología Celular
, Rodríguez Gallardo, Sofía
, Okazaki, Kouta
, European Commission (EC). Fondo Europeo de Desarrollo Regional (FEDER)
, Muñiz Guinea, Manuel
, López Martín, Sergio
, Aguilera Romero, María Auxiliadora
, Ministerio de Ciencia, Innovación y Universidades (MICINN). España
, Ikeda, Atsuko
, Nakano, Miyako
, Japan Society for the Promotion of Science
, Sabido Bozo, Susana
in
Analysis
/ Biology and Life Sciences
/ Cell membranes
/ Cell surface
/ Electrophoresis
/ Endoplasmic reticulum
/ Enzymes
/ Eukaryotes
/ Glycan
/ Glycolipids
/ Glycosylphosphatidylinositol
/ Ions
/ Lab Protocol
/ Mass spectrometry
/ Mass spectroscopy
/ Peptides
/ Plasma
/ Polysaccharides
/ Post-translation
/ Proteins
/ Research and Analysis Methods
/ Scientific imaging
/ Structural analysis
/ Structure
/ Trypsin
/ Yeast
/ Yeasts
2021
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Structural analysis of the GPI glycan
by
Funato, Kouichi
, Cortés Gómez, Alejandro
, Universidad de Sevilla. Departamento de Biología Celular
, Rodríguez Gallardo, Sofía
, Okazaki, Kouta
, European Commission (EC). Fondo Europeo de Desarrollo Regional (FEDER)
, Muñiz Guinea, Manuel
, López Martín, Sergio
, Aguilera Romero, María Auxiliadora
, Ministerio de Ciencia, Innovación y Universidades (MICINN). España
, Ikeda, Atsuko
, Nakano, Miyako
, Japan Society for the Promotion of Science
, Sabido Bozo, Susana
in
Analysis
/ Biology and Life Sciences
/ Cell membranes
/ Cell surface
/ Electrophoresis
/ Endoplasmic reticulum
/ Enzymes
/ Eukaryotes
/ Glycan
/ Glycolipids
/ Glycosylphosphatidylinositol
/ Ions
/ Lab Protocol
/ Mass spectrometry
/ Mass spectroscopy
/ Peptides
/ Plasma
/ Polysaccharides
/ Post-translation
/ Proteins
/ Research and Analysis Methods
/ Scientific imaging
/ Structural analysis
/ Structure
/ Trypsin
/ Yeast
/ Yeasts
2021
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Journal Article
Structural analysis of the GPI glycan
2021
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Overview
Glycosylphosphatidylinositol (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.
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