Exploring the Nicotinic Acetylcholine Receptor-associated Proteome with iTRAQ and Transgenic Mice

Neuronal nicotinic acetylcholine receptors (nAChRs) containing α4 and β2 subunits are the principal receptors in the mammalian central nervous system that bind nicotine with high affinity. These nAChRs are involved in nicotine dependence, mood disorders, neurodegeneration and neuroprotection. Howeve...

Descripción completa

Detalles Bibliográficos
Autores principales: McClure-Begley, Tristan D., Stone, Kathy L., Marks, Michael J., Grady, Sharon R., Colangelo, Christopher M., Lindstrom, Jon M., Picciotto, Marina R.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2013
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3806329/
https://www.ncbi.nlm.nih.gov/pubmed/23891776
http://dx.doi.org/10.1016/j.gpb.2013.05.005
_version_ 1782288362034429952
author McClure-Begley, Tristan D.
Stone, Kathy L.
Marks, Michael J.
Grady, Sharon R.
Colangelo, Christopher M.
Lindstrom, Jon M.
Picciotto, Marina R.
author_facet McClure-Begley, Tristan D.
Stone, Kathy L.
Marks, Michael J.
Grady, Sharon R.
Colangelo, Christopher M.
Lindstrom, Jon M.
Picciotto, Marina R.
author_sort McClure-Begley, Tristan D.
collection PubMed
description Neuronal nicotinic acetylcholine receptors (nAChRs) containing α4 and β2 subunits are the principal receptors in the mammalian central nervous system that bind nicotine with high affinity. These nAChRs are involved in nicotine dependence, mood disorders, neurodegeneration and neuroprotection. However, our understanding of the interactions between α4β2-containing (α4β2(∗)) nAChRs and other proteins remains limited. In this study, we identified proteins that interact with α4β2(∗) nAChRs in a genedose dependent pattern by immunopurifying β2(∗) nAChRs from mice that differ in α4 and β2 subunit expression and performing proteomic analysis using isobaric tags for relative and absolute quantitation (iTRAQ). Reduced expression of either the α4 or the β2 subunit results in a correlated decline in the expression of a number of putative interacting proteins. We identified 208 proteins co-immunoprecipitated with these nAChRs. Furthermore, stratified linear regression analysis indicated that levels of 17 proteins was correlated significantly with expression of α4β2 nAChRs, including proteins involved in cytoskeletal rearrangement and calcium signaling. These findings represent the first application of quantitative proteomics to produce a β2(∗) nAChR interactome and describe a novel technique used to discover potential targets for pharmacological manipulation of α4β2 nAChRs and their downstream signaling mechanisms.
format Online
Article
Text
id pubmed-3806329
institution National Center for Biotechnology Information
language English
publishDate 2013
publisher Elsevier
record_format MEDLINE/PubMed
spelling pubmed-38063292013-10-23 Exploring the Nicotinic Acetylcholine Receptor-associated Proteome with iTRAQ and Transgenic Mice McClure-Begley, Tristan D. Stone, Kathy L. Marks, Michael J. Grady, Sharon R. Colangelo, Christopher M. Lindstrom, Jon M. Picciotto, Marina R. Genomics Proteomics Bioinformatics Original Research Neuronal nicotinic acetylcholine receptors (nAChRs) containing α4 and β2 subunits are the principal receptors in the mammalian central nervous system that bind nicotine with high affinity. These nAChRs are involved in nicotine dependence, mood disorders, neurodegeneration and neuroprotection. However, our understanding of the interactions between α4β2-containing (α4β2(∗)) nAChRs and other proteins remains limited. In this study, we identified proteins that interact with α4β2(∗) nAChRs in a genedose dependent pattern by immunopurifying β2(∗) nAChRs from mice that differ in α4 and β2 subunit expression and performing proteomic analysis using isobaric tags for relative and absolute quantitation (iTRAQ). Reduced expression of either the α4 or the β2 subunit results in a correlated decline in the expression of a number of putative interacting proteins. We identified 208 proteins co-immunoprecipitated with these nAChRs. Furthermore, stratified linear regression analysis indicated that levels of 17 proteins was correlated significantly with expression of α4β2 nAChRs, including proteins involved in cytoskeletal rearrangement and calcium signaling. These findings represent the first application of quantitative proteomics to produce a β2(∗) nAChR interactome and describe a novel technique used to discover potential targets for pharmacological manipulation of α4β2 nAChRs and their downstream signaling mechanisms. Elsevier 2013-08 2013-07-25 /pmc/articles/PMC3806329/ /pubmed/23891776 http://dx.doi.org/10.1016/j.gpb.2013.05.005 Text en © 2013 Beijing Institute of Genomics, Chinese Academy of Sciences and Genetics Society of China. Production and hosting by Elsevier B.V. All rights reserved. http://creativecommons.org/licenses/by-nc-sa/3.0/ This is an open access article under the CC BY-NC-SA license (http://creativecommons.org/licenses/by-nc-sa/3.0/).
spellingShingle Original Research
McClure-Begley, Tristan D.
Stone, Kathy L.
Marks, Michael J.
Grady, Sharon R.
Colangelo, Christopher M.
Lindstrom, Jon M.
Picciotto, Marina R.
Exploring the Nicotinic Acetylcholine Receptor-associated Proteome with iTRAQ and Transgenic Mice
title Exploring the Nicotinic Acetylcholine Receptor-associated Proteome with iTRAQ and Transgenic Mice
title_full Exploring the Nicotinic Acetylcholine Receptor-associated Proteome with iTRAQ and Transgenic Mice
title_fullStr Exploring the Nicotinic Acetylcholine Receptor-associated Proteome with iTRAQ and Transgenic Mice
title_full_unstemmed Exploring the Nicotinic Acetylcholine Receptor-associated Proteome with iTRAQ and Transgenic Mice
title_short Exploring the Nicotinic Acetylcholine Receptor-associated Proteome with iTRAQ and Transgenic Mice
title_sort exploring the nicotinic acetylcholine receptor-associated proteome with itraq and transgenic mice
topic Original Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3806329/
https://www.ncbi.nlm.nih.gov/pubmed/23891776
http://dx.doi.org/10.1016/j.gpb.2013.05.005
work_keys_str_mv AT mcclurebegleytristand exploringthenicotinicacetylcholinereceptorassociatedproteomewithitraqandtransgenicmice
AT stonekathyl exploringthenicotinicacetylcholinereceptorassociatedproteomewithitraqandtransgenicmice
AT marksmichaelj exploringthenicotinicacetylcholinereceptorassociatedproteomewithitraqandtransgenicmice
AT gradysharonr exploringthenicotinicacetylcholinereceptorassociatedproteomewithitraqandtransgenicmice
AT colangelochristopherm exploringthenicotinicacetylcholinereceptorassociatedproteomewithitraqandtransgenicmice
AT lindstromjonm exploringthenicotinicacetylcholinereceptorassociatedproteomewithitraqandtransgenicmice
AT picciottomarinar exploringthenicotinicacetylcholinereceptorassociatedproteomewithitraqandtransgenicmice