Structural analysis of the GPI glycan.

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 rol...

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Autores principales:	Miyako Nakano, Susana Sabido-Bozo, Kouta Okazaki, Auxiliadora Aguilera-Romero, Sofia Rodriguez-Gallardo, Alejandro Cortes-Gomez, Sergio Lopez, Atsuko Ikeda, Kouichi Funato, Manuel Muñiz
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/d314bcf6489d4e8b90bacebc982dad0d
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spelling	oai:doaj.org-article:d314bcf6489d4e8b90bacebc982dad0d2021-12-02T20:14:34ZStructural analysis of the GPI glycan.1932-620310.1371/journal.pone.0257435https://doaj.org/article/d314bcf6489d4e8b90bacebc982dad0d2021-01-01T00:00:00Zhttps://doi.org/10.1371/journal.pone.0257435https://doaj.org/toc/1932-6203Glycosylphosphatidylinositol (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.Miyako NakanoSusana Sabido-BozoKouta OkazakiAuxiliadora Aguilera-RomeroSofia Rodriguez-GallardoAlejandro Cortes-GomezSergio LopezAtsuko IkedaKouichi FunatoManuel MuñizPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 16, Iss 9, p e0257435 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Miyako Nakano Susana Sabido-Bozo Kouta Okazaki Auxiliadora Aguilera-Romero Sofia Rodriguez-Gallardo Alejandro Cortes-Gomez Sergio Lopez Atsuko Ikeda Kouichi Funato Manuel Muñiz Structural analysis of the GPI glycan.
description	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.
format	article
author	Miyako Nakano Susana Sabido-Bozo Kouta Okazaki Auxiliadora Aguilera-Romero Sofia Rodriguez-Gallardo Alejandro Cortes-Gomez Sergio Lopez Atsuko Ikeda Kouichi Funato Manuel Muñiz
author_facet	Miyako Nakano Susana Sabido-Bozo Kouta Okazaki Auxiliadora Aguilera-Romero Sofia Rodriguez-Gallardo Alejandro Cortes-Gomez Sergio Lopez Atsuko Ikeda Kouichi Funato Manuel Muñiz
author_sort	Miyako Nakano
title	Structural analysis of the GPI glycan.
title_short	Structural analysis of the GPI glycan.
title_full	Structural analysis of the GPI glycan.
title_fullStr	Structural analysis of the GPI glycan.
title_full_unstemmed	Structural analysis of the GPI glycan.
title_sort	structural analysis of the gpi glycan.
publisher	Public Library of Science (PLoS)
publishDate	2021
url	https://doaj.org/article/d314bcf6489d4e8b90bacebc982dad0d
work_keys_str_mv	AT miyakonakano structuralanalysisofthegpiglycan AT susanasabidobozo structuralanalysisofthegpiglycan AT koutaokazaki structuralanalysisofthegpiglycan AT auxiliadoraaguileraromero structuralanalysisofthegpiglycan AT sofiarodriguezgallardo structuralanalysisofthegpiglycan AT alejandrocortesgomez structuralanalysisofthegpiglycan AT sergiolopez structuralanalysisofthegpiglycan AT atsukoikeda structuralanalysisofthegpiglycan AT kouichifunato structuralanalysisofthegpiglycan AT manuelmuniz structuralanalysisofthegpiglycan
_version_	1718374672867786752

Structural analysis of the GPI glycan.

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