Structural analysis of the GPI glycan
Sabido Bozo, Susana
Aguilera Romero, María Auxiliadora
Rodríguez Gallardo, Sofía
Cortés Gómez, Alejandro
López Martín, Sergio
Muñiz Guinea, Manuel
|Department||Universidad de Sevilla. Departamento de Biología Celular|
|Abstract||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) ...
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.
|Funding agencies||Ministerio de Ciencia, Innovación y Universidades (MICINN). España
European Commission (EC). Fondo Europeo de Desarrollo Regional (FEDER)
Japan Society for the Promotion of Science
|Citation||Nakano, M., Sabido Bozo, S., Okazaki, K., Aguilera Romero, M.A., Rodríguez Gallardo, S., Cortés Gómez, A.,...,Muñiz Guinea, M. (2021). Structural analysis of the GPI glycan. PLOS ONE, 16 (9), e0257435.|