RT Journal Article
T1 Determination of the lipid composition of the GPI anchor
A1 Aguilera-Romero, Auxiliadora
A1 Sabido-Bozo, Susana
A1 Lopez, Sergio
A1 Cortes-Gomez, Alejandro
A1 Rodriguez-Gallardo, Sofia
A1 Perez-Linero, Ana Maria
A1 Riezman, Isabelle
A1 Riezman, Howard
A1 Muñiz, Manuel
K1 Lipids
K1 Glycosylphosphatidylinositols
K1 Membranes
K1 Ceramides
K1 Genes, reporter
K1 Lípidos
K1 Glicosilfosfatidilinositoles
K1 Membranas
K1 Ceramidas
K1 Genes informadores
AB In 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.
PB Public Library of Science
YR 2021
FD 2021-08-13
LK http://hdl.handle.net/10668/4314
UL http://hdl.handle.net/10668/4314
LA en
NO Aguilera-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):e0256184
DS RISalud
RD Apr 11, 2025