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Exploring the Nicotinic Acetylcholine Receptor-associated Proteome with iTRAQ and Transgenic Mice 被引量:1
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作者 Tristan D. McClure-Begley Kathy L. Stone +4 位作者 Michael J. Marks Sharon R. Grady Christopher M. Colangelo Jon M. Lindstrom Marina R. Picciotto 《Genomics, Proteomics & Bioinformatics》 SCIE CAS CSCD 2013年第4期207-218,共12页
Neuronal nicotinic acetylcholine receptors (nAChRs) containing Gt4 and 132 subunits are the principal receptors in the mammalian central nervous system that bind nicotine with high affin- ity. These nAChRs are invol... Neuronal nicotinic acetylcholine receptors (nAChRs) containing Gt4 and 132 subunits are the principal receptors in the mammalian central nervous system that bind nicotine with high affin- ity. These nAChRs are involved in nicotine dependence, mood disorders, neurodegeneration and neuroprotection. However, our understanding of the interactions between a4β2-containing (a4β2) nAChRs and other proteins remains limited. In this study, we identified proteins that inter- act with ct4β2 nAChRs in a gene-dose dependent pattern by immunopurifying β2 nAChRs from mice that differ in ct4 and β2 subunit expression and performing proteomic analysis using isobaric tags for relative and absolute quantitation (iTRAQ). Reduced expression of either the a4 or the β2 subunit results in a correlated decline in the expression of a number of putative interacting proteins. We identified 208 proteins co-imrnunoprecipitated with these nAChRs. Furthermore, stratified lin- ear regression analysis indicated that levels of 17 proteins was correlated significantly with expres- sion of at4β2 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 pharma- cological manipulation of a4β2 nAChRs and their downstream signaling mechanisms. 展开更多
关键词 Nicotinic receptor Affinity purification Quantitative proteomics Transgenic mouse
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