Expressed glycosylphosphatidylinositol-anchored horseradish peroxidase identifies co-clustering molecules in individual lipid raft domains.

Lipid rafts that are enriched in glycosylphosphatidylinositol (GPI)-anchored proteins serve as a platform for important biological events. To elucidate the molecular mechanisms of these events, identification of co-clustering molecules in individual raft domains is required. Here we describe an appr...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Arisa Miyagawa-Yamaguchi, Norihiro Kotani, Koichi Honke
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2014
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/9990bc6065794f9183324888b531aed5
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:9990bc6065794f9183324888b531aed5
record_format	dspace
spelling	oai:doaj.org-article:9990bc6065794f9183324888b531aed52021-11-18T08:26:04ZExpressed glycosylphosphatidylinositol-anchored horseradish peroxidase identifies co-clustering molecules in individual lipid raft domains.1932-620310.1371/journal.pone.0093054https://doaj.org/article/9990bc6065794f9183324888b531aed52014-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24671047/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Lipid rafts that are enriched in glycosylphosphatidylinositol (GPI)-anchored proteins serve as a platform for important biological events. To elucidate the molecular mechanisms of these events, identification of co-clustering molecules in individual raft domains is required. Here we describe an approach to this issue using the recently developed method termed enzyme-mediated activation of radical source (EMARS), by which molecules in the vicinity within 300 nm from horseradish peroxidase (HRP) set on the probed molecule are labeled. GPI-anchored HRP fusion proteins (HRP-GPIs), in which the GPI attachment signals derived from human decay accelerating factor and Thy-1 were separately connected to the C-terminus of HRP, were expressed in HeLa S3 cells, and the EMARS reaction was catalyzed by these expressed HRP-GPIs under a living condition. As a result, these different HRP-GPIs had differences in glycosylation and localization and formed distinct clusters. This novel approach distinguished molecular clusters associated with individual GPI-anchored proteins, suggesting that it can identify co-clustering molecules in individual raft domains.Arisa Miyagawa-YamaguchiNorihiro KotaniKoichi HonkePublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 9, Iss 3, p e93054 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Arisa Miyagawa-Yamaguchi Norihiro Kotani Koichi Honke Expressed glycosylphosphatidylinositol-anchored horseradish peroxidase identifies co-clustering molecules in individual lipid raft domains.
description	Lipid rafts that are enriched in glycosylphosphatidylinositol (GPI)-anchored proteins serve as a platform for important biological events. To elucidate the molecular mechanisms of these events, identification of co-clustering molecules in individual raft domains is required. Here we describe an approach to this issue using the recently developed method termed enzyme-mediated activation of radical source (EMARS), by which molecules in the vicinity within 300 nm from horseradish peroxidase (HRP) set on the probed molecule are labeled. GPI-anchored HRP fusion proteins (HRP-GPIs), in which the GPI attachment signals derived from human decay accelerating factor and Thy-1 were separately connected to the C-terminus of HRP, were expressed in HeLa S3 cells, and the EMARS reaction was catalyzed by these expressed HRP-GPIs under a living condition. As a result, these different HRP-GPIs had differences in glycosylation and localization and formed distinct clusters. This novel approach distinguished molecular clusters associated with individual GPI-anchored proteins, suggesting that it can identify co-clustering molecules in individual raft domains.
format	article
author	Arisa Miyagawa-Yamaguchi Norihiro Kotani Koichi Honke
author_facet	Arisa Miyagawa-Yamaguchi Norihiro Kotani Koichi Honke
author_sort	Arisa Miyagawa-Yamaguchi
title	Expressed glycosylphosphatidylinositol-anchored horseradish peroxidase identifies co-clustering molecules in individual lipid raft domains.
title_short	Expressed glycosylphosphatidylinositol-anchored horseradish peroxidase identifies co-clustering molecules in individual lipid raft domains.
title_full	Expressed glycosylphosphatidylinositol-anchored horseradish peroxidase identifies co-clustering molecules in individual lipid raft domains.
title_fullStr	Expressed glycosylphosphatidylinositol-anchored horseradish peroxidase identifies co-clustering molecules in individual lipid raft domains.
title_full_unstemmed	Expressed glycosylphosphatidylinositol-anchored horseradish peroxidase identifies co-clustering molecules in individual lipid raft domains.
title_sort	expressed glycosylphosphatidylinositol-anchored horseradish peroxidase identifies co-clustering molecules in individual lipid raft domains.
publisher	Public Library of Science (PLoS)
publishDate	2014
url	https://doaj.org/article/9990bc6065794f9183324888b531aed5
work_keys_str_mv	AT arisamiyagawayamaguchi expressedglycosylphosphatidylinositolanchoredhorseradishperoxidaseidentifiescoclusteringmoleculesinindividuallipidraftdomains AT norihirokotani expressedglycosylphosphatidylinositolanchoredhorseradishperoxidaseidentifiescoclusteringmoleculesinindividuallipidraftdomains AT koichihonke expressedglycosylphosphatidylinositolanchoredhorseradishperoxidaseidentifiescoclusteringmoleculesinindividuallipidraftdomains
_version_	1718421819102330880

Expressed glycosylphosphatidylinositol-anchored horseradish peroxidase identifies co-clustering molecules in individual lipid raft domains.

Ejemplares similares