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A Redox-Sensitive Luciferase Assay for Determining the Localization and Topology of Endoplasmic Reticulum Proteins

Correct localization and transmembrane topology are crucial for the proteins residing and functioning in the endoplasmic reticulum (ER). We have developed a rapid and convenient assay, based on the redox-sensitive luciferase from Gaussia princeps (Gluc) and green fluorescence protein (GFP), to deter...

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Autores principales: Li, Hai-Yin, Zheng, Xue-Ming, Che, Mei-Xia, Hu, Hong-Yu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3329452/
https://www.ncbi.nlm.nih.gov/pubmed/22530060
http://dx.doi.org/10.1371/journal.pone.0035628
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author Li, Hai-Yin
Zheng, Xue-Ming
Che, Mei-Xia
Hu, Hong-Yu
author_facet Li, Hai-Yin
Zheng, Xue-Ming
Che, Mei-Xia
Hu, Hong-Yu
author_sort Li, Hai-Yin
collection PubMed
description Correct localization and transmembrane topology are crucial for the proteins residing and functioning in the endoplasmic reticulum (ER). We have developed a rapid and convenient assay, based on the redox-sensitive luciferase from Gaussia princeps (Gluc) and green fluorescence protein (GFP), to determine the localization or topology of ER proteins. Using the tandem Gluc-GFP reporter fused to different positions of a target protein, we successfully characterized the topologies of two ER transmembrane proteins Herp and HRD1 that are involved in the ER quality control system. This assay method may also be applicable to the proteins in secretory pathway, plasma membrane, and other compartments of cells.
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spelling pubmed-33294522012-04-23 A Redox-Sensitive Luciferase Assay for Determining the Localization and Topology of Endoplasmic Reticulum Proteins Li, Hai-Yin Zheng, Xue-Ming Che, Mei-Xia Hu, Hong-Yu PLoS One Research Article Correct localization and transmembrane topology are crucial for the proteins residing and functioning in the endoplasmic reticulum (ER). We have developed a rapid and convenient assay, based on the redox-sensitive luciferase from Gaussia princeps (Gluc) and green fluorescence protein (GFP), to determine the localization or topology of ER proteins. Using the tandem Gluc-GFP reporter fused to different positions of a target protein, we successfully characterized the topologies of two ER transmembrane proteins Herp and HRD1 that are involved in the ER quality control system. This assay method may also be applicable to the proteins in secretory pathway, plasma membrane, and other compartments of cells. Public Library of Science 2012-04-18 /pmc/articles/PMC3329452/ /pubmed/22530060 http://dx.doi.org/10.1371/journal.pone.0035628 Text en Li et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Li, Hai-Yin
Zheng, Xue-Ming
Che, Mei-Xia
Hu, Hong-Yu
A Redox-Sensitive Luciferase Assay for Determining the Localization and Topology of Endoplasmic Reticulum Proteins
title A Redox-Sensitive Luciferase Assay for Determining the Localization and Topology of Endoplasmic Reticulum Proteins
title_full A Redox-Sensitive Luciferase Assay for Determining the Localization and Topology of Endoplasmic Reticulum Proteins
title_fullStr A Redox-Sensitive Luciferase Assay for Determining the Localization and Topology of Endoplasmic Reticulum Proteins
title_full_unstemmed A Redox-Sensitive Luciferase Assay for Determining the Localization and Topology of Endoplasmic Reticulum Proteins
title_short A Redox-Sensitive Luciferase Assay for Determining the Localization and Topology of Endoplasmic Reticulum Proteins
title_sort redox-sensitive luciferase assay for determining the localization and topology of endoplasmic reticulum proteins
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3329452/
https://www.ncbi.nlm.nih.gov/pubmed/22530060
http://dx.doi.org/10.1371/journal.pone.0035628
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