RT Journal Article
T1 Structural analysis of the GPI glycan.
A1 Nakano, Miyako
A1 Sabido-Bozo, Susana
A1 Okazaki, Kouta
A1 Aguilera-Romero, Auxiliadora
A1 Rodriguez-Gallardo, Sofia
A1 Cortes-Gomez, Alejandro
A1 Lopez, Sergio
A1 Ikeda, Atsuko
A1 Funato, Kouichi
A1 Muñiz, Manuel
AB 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.
YR 2021
FD 2021-09-16
LK https://hdl.handle.net/10668/27874
UL https://hdl.handle.net/10668/27874
LA en
DS RISalud
RD Apr 11, 2025