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Snorkel: An Epitope Tagging System for Measuring the Surface Expression of Membrane Proteins
Tags are widely used to monitor a protein’s expression level, interactions, protein trafficking, and localization. Membrane proteins are often tagged in their extracellular domains to allow discrimination between protein in the plasma membrane from that in internal pools. Multipass membrane proteins...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2013
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3759426/ https://www.ncbi.nlm.nih.gov/pubmed/24023844 http://dx.doi.org/10.1371/journal.pone.0073255 |
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author | Brown, Michael Stafford, Lewis J. Onisk, Dale Joaquim, Tony Tobb, Alhagie Goldman, Larissa Fancy, David Stave, James Chambers, Ross |
author_facet | Brown, Michael Stafford, Lewis J. Onisk, Dale Joaquim, Tony Tobb, Alhagie Goldman, Larissa Fancy, David Stave, James Chambers, Ross |
author_sort | Brown, Michael |
collection | PubMed |
description | Tags are widely used to monitor a protein’s expression level, interactions, protein trafficking, and localization. Membrane proteins are often tagged in their extracellular domains to allow discrimination between protein in the plasma membrane from that in internal pools. Multipass membrane proteins offer special challenges for inserting a tag since the extracellular regions are often composed of small loops and thus inserting an epitope tag risks perturbing the structure, function, or location of the membrane protein. We have developed a novel tagging system called snorkel where a transmembrane domain followed by a tag is appended to the cytoplasmic C-terminus of the membrane protein. In this way the tag is displayed extracellularly, but structurally separate from the membrane protein. We have tested the snorkel tag system on a diverse panel of membrane proteins including GPCRs and ion channels and demonstrated that it reliably allows for monitoring of the surface expression. |
format | Online Article Text |
id | pubmed-3759426 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-37594262013-09-10 Snorkel: An Epitope Tagging System for Measuring the Surface Expression of Membrane Proteins Brown, Michael Stafford, Lewis J. Onisk, Dale Joaquim, Tony Tobb, Alhagie Goldman, Larissa Fancy, David Stave, James Chambers, Ross PLoS One Research Article Tags are widely used to monitor a protein’s expression level, interactions, protein trafficking, and localization. Membrane proteins are often tagged in their extracellular domains to allow discrimination between protein in the plasma membrane from that in internal pools. Multipass membrane proteins offer special challenges for inserting a tag since the extracellular regions are often composed of small loops and thus inserting an epitope tag risks perturbing the structure, function, or location of the membrane protein. We have developed a novel tagging system called snorkel where a transmembrane domain followed by a tag is appended to the cytoplasmic C-terminus of the membrane protein. In this way the tag is displayed extracellularly, but structurally separate from the membrane protein. We have tested the snorkel tag system on a diverse panel of membrane proteins including GPCRs and ion channels and demonstrated that it reliably allows for monitoring of the surface expression. Public Library of Science 2013-09-02 /pmc/articles/PMC3759426/ /pubmed/24023844 http://dx.doi.org/10.1371/journal.pone.0073255 Text en © 2013 Brown 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 Brown, Michael Stafford, Lewis J. Onisk, Dale Joaquim, Tony Tobb, Alhagie Goldman, Larissa Fancy, David Stave, James Chambers, Ross Snorkel: An Epitope Tagging System for Measuring the Surface Expression of Membrane Proteins |
title | Snorkel: An Epitope Tagging System for Measuring the Surface Expression of Membrane Proteins |
title_full | Snorkel: An Epitope Tagging System for Measuring the Surface Expression of Membrane Proteins |
title_fullStr | Snorkel: An Epitope Tagging System for Measuring the Surface Expression of Membrane Proteins |
title_full_unstemmed | Snorkel: An Epitope Tagging System for Measuring the Surface Expression of Membrane Proteins |
title_short | Snorkel: An Epitope Tagging System for Measuring the Surface Expression of Membrane Proteins |
title_sort | snorkel: an epitope tagging system for measuring the surface expression of membrane proteins |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3759426/ https://www.ncbi.nlm.nih.gov/pubmed/24023844 http://dx.doi.org/10.1371/journal.pone.0073255 |
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