Targeted tandem affinity purification of PSD-95 recovers core postsynaptic complexes and schizophrenia susceptibility proteins.
Targeted tandem affinity purification of PSD-95 recovers core postsynaptic complexes and schizophrenia susceptibility proteins.
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DOI：
10.1038/msb.2009.27
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发表时间：
2009
影响因子：
9.9
通讯作者：
Grant, Seth G. N.
中科院分区：
文献类型：
--
作者：
Fernandez, Esperanza;Collins, Mark O.;Uren, Rachel T.;Kopanitsa, Maksym V.;Komiyama, Noboru H.;Croning, Mike D. R.;Zografos, Lysimachos;Armstrong, J. Douglas;Choudhary, Jyoti S.;Grant, Seth G. N.
关键词：
哺乳动物蛋白质组的分子复杂性需要新的方法来绘制多蛋白复合物的组织。在这里，我们结合小鼠遗传学和蛋白质组学来表征突触蛋白复合物和相互作用网络。利用基因靶向技术将新的串联亲和纯化（TAP）标签融合到小鼠PSD-95的羧基端。纯合子小鼠在PSD-95表达、亚细胞定位和突触电生理功能方面均未发现异常。通过质谱分析在天然条件下纯化的多蛋白复合物，定义了已知的和新的相互作用物：118种包含突触关键功能成分的蛋白质，包括谷氨酸受体、K+通道、支架和信号蛋白。蛋白质相互作用的网络聚类产生了五个相连的簇，其中两个簇包含所有主要的嗜离子性谷氨酸受体，一个簇包含电压依赖的K+通道。与人类疾病相关的簇的注释显示，多种疾病映射到网络中，在谷氨酸受体簇中精神分裂症具有显着相关性。这种靶向TAP标记策略通常适用于哺乳动物蛋白质组学和疾病的系统生物学方法。
The molecular complexity of mammalian proteomes demands new methods for mapping the organization of multiprotein complexes. Here, we combine mouse genetics and proteomics to characterize synapse protein complexes and interaction networks. New tandem affinity purification (TAP) tags were fused to the carboxyl terminus of PSD-95 using gene targeting in mice. Homozygous mice showed no detectable abnormalities in PSD-95 expression, subcellular localization or synaptic electrophysiological function. Analysis of multiprotein complexes purified under native conditions by mass spectrometry defined known and new interactors: 118 proteins comprising crucial functional components of synapses, including glutamate receptors, K+ channels, scaffolding and signaling proteins, were recovered. Network clustering of protein interactions generated five connected clusters, with two clusters containing all the major ionotropic glutamate receptors and one cluster with voltage-dependent K+ channels. Annotation of clusters with human disease associations revealed that multiple disorders map to the network, with a significant correlation of schizophrenia within the glutamate receptor clusters. This targeted TAP tagging strategy is generally applicable to mammalian proteomics and systems biology approaches to disease.
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DOI：
10.1523/jneurosci.4457-06.2007
发表时间：
2007-03-07
期刊：
The Journal of neuroscience : the official journal of the Society for Neuroscience
影响因子：
--
作者：
Cuthbert PC;Stanford LE;Coba MP;Ainge JA;Fink AE;Opazo P;Delgado JY;Komiyama NH;O'Dell TJ;Grant SG
通讯作者：
Grant SG
影响因子：
4.8
作者：
Collins, MO;Yu, L;Grant, SGN
通讯作者：
Grant, SGN
影响因子：
16.2
作者：
Chowdhury, Shoaib;Shepherd, Jason D.;Worley, Paul F.
通讯作者：
Worley, Paul F.
影响因子：
9.2
作者：
Garry, EM;Moss, A;Fleetwood-Walker, SM
通讯作者：
Fleetwood-Walker, SM
影响因子：
4.7
作者：
Husi, H;Grant, SGN
通讯作者：
Grant, SGN