Proteomic analysis of an alpha7 nicotinic acetylcholine receptor interactome.
Proteomic analysis of an alpha7 nicotinic acetylcholine receptor interactome.
复制标题
DOI：
10.1021/pr800731z
复制
发表时间：
2009-04
影响因子：
4.4
通讯作者：
Hawrot E
中科院分区：
文献类型：
--
作者：
Paulo JA;Brucker WJ;Hawrot E
α7烟碱乙酰胆碱受体（nictinic acetylcholine receptor, nAChR）是哺乳动物脑内主要的高亲和力α-班加罗毒素结合蛋白。采用亲和固定和选择性洗脱的方法，从小鼠脑组织中分离出对碳甾醇敏感的α-班加罗毒素结合复合物，并用SDS-PAGE进行分离。将细分的凝胶通道片段中的蛋白质进行胰酶消化，并通过标准质谱法分析所得肽。我们在野生型样品中鉴定了55种蛋白质，这些蛋白质在α7 nAChR敲除小鼠的类似脑样品中不存在，这些样品已经以平行方式处理。这55种蛋白中的许多都是α7 nAChR相互作用伙伴的新蛋白质组候选蛋白，并且许多与可能涉及α7在中枢神经系统中的功能的多种信号通路相关。新发现的潜在蛋白质相互作用，以及我们介绍的α-虫毒结合蛋白复合物的一般方法，为许多有趣的后续研究提供了一个新的平台，旨在阐明神经元α7 nachr的生理作用。
The α7 nicotinic acetylcholine receptor (nAChR) is well established as the principal high-affinity α-bungarotoxin-binding protein in the mammalian brain. We isolated carbachol-sensitive α-bungarotoxin-binding complexes from total mouse brain tissue by affinity immobilization followed by selective elution, and these proteins were fractionated by SDS-PAGE. The proteins in subdivided gel lane segments were tryptically digested, and the resulting peptides were analyzed by standard mass spectrometry. We identified 55 proteins in wild-type samples that were not present in comparable brain samples from α7 nAChR knockout mice that had been processed in a parallel fashion. Many of these 55 proteins are novel proteomic candidates for interaction partners of the α7 nAChR, and many are associated with multiple signaling pathways that may be implicated in α7 function in the central nervous system. The newly identified potential protein interactions, together with the general methodology that we introduce for α-bungarotoxin-binding protein complexes, form a new platform for many interesting follow-up studies aimed at elucidating the physiological role of neuronal α7 nAChRs.
登录
查看更多内容
影响因子：
3.5
作者：
Ango, F;Robbe, D;Fagni, L
通讯作者：
Fagni, L
影响因子：
5.3
作者：
Fischer, H;Liu, DM;Adams, DJ
通讯作者：
Adams, DJ
影响因子：
9.3
作者：
De Simone R;Ajmone-Cat MA;Carnevale D;Minghetti L
通讯作者：
Minghetti L
影响因子：
16.2
作者：
FUNTE, LR;HAYDON, PG
通讯作者：
HAYDON, PG
影响因子：
3.5
作者：
Furnish, EJ;Zhou, W;Schmidt, CE
通讯作者：
Schmidt, CE