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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...

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Detalles Bibliográficos
Autores principales: Brown, Michael, Stafford, Lewis J., Onisk, Dale, Joaquim, Tony, Tobb, Alhagie, Goldman, Larissa, Fancy, David, Stave, James, Chambers, Ross
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2013
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.
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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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