Affinity-purification mass spectrometry (AP-MS) of serine/threonine phosphatases
Affinity-purification mass spectrometry (AP-MS) of serine/threonine phosphatases
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DOI：
10.1016/j.ymeth.2007.02.018
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发表时间：
2007-07-01
期刊：
影响因子：
4.8
通讯作者：
Gingras, Anne-Claude
中科院分区：
文献类型：
--
作者：
Chen, Ginny I.;Gingras, Anne-Claude
丝氨酸/苏氨酸磷酸酶与调节蛋白的关联是其特异性的关键组成部分，并且这些结合伴侣的鉴定对于理解磷酸酶的功能和调节至关重要。亲和纯化/质谱（AP-MS）方法已经并将继续在确定这些相互作用。在这里，我们回顾了AP-MS的一般原理，并提出了两个亲和纯化协议兼容随后的质谱，即FLAG纯化，和串联亲和纯化（TAP）。我们已经成功地使用这些协议的识别PP 2A，PP 4和PP 6的结合伙伴，他们应该服从于其他磷酸酶的相互作用的分析。(c)2007爱思唯尔公司All rights reserved.
Association of serine/threonine phosphatases with regulatory proteins is a key component of their specificity, and the identification of these binding partners is critical to understanding phosphatases function and regulation. Affinity-purification/mass spectrometry (AP-MS) approaches have been and continue to be instrumental in identifying these interactors. Here, we review the general principles of AP-MS, and present two affinity-purification protocols compatible with subsequent mass spectrometry, namely FLAG purification, and the tandem affinity purification (TAP). We have successfully used these protocols for the identification of binding partners for PP2A, PP4 and PP6, and they should be amenable to the analysis of interactors for other phosphatases. (c) 2007 Elsevier Inc. All rights reserved.