A proteomic study of SUMO-2 target proteins
A proteomic study of SUMO-2 target proteins
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DOI：
10.1074/jbc.m404201200
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发表时间：
2004-08-06
影响因子：
4.8
通讯作者：
Lamond, AI
中科院分区：
文献类型：
--
作者：
Vertegaal, ACO;Ogg, SC;Lamond, AI
脊椎动物中的SUMO家族包括至少三种不同的蛋白（SUMO-1、-2和-3），它们作为翻译后修饰添加到靶蛋白上。相当数量的SUMO-1靶蛋白已被确定，但对SUMO-2知之甚少。建立了表达His(6)标记的SUMO-2的稳定HeLa细胞系，并用于标记和纯化新的内源性SUMO-2靶蛋白。SUMO-2的标记形式是功能性的，主要定位于细胞核。他的(6)标记的SUMO-2共轭物从核部分亲和纯化，并通过质谱鉴定。八个新的潜在SUMO-2靶蛋白被至少两个肽鉴定。其中三种蛋白，SART1、异质核核糖核蛋白（RNP） M和U5小核RNP 200-kDa解旋酶，在RNA代谢中发挥作用。SART1和异质核RNP M都被证明是真正的SUMO目标，证实了该方法的有效性。
The SUMO family in vertebrates includes at least three distinct proteins ( SUMO-1, -2, and -3) that are added as post-translational modifications to target proteins. A considerable number of SUMO-1 target proteins have been identified, but little is known about SUMO-2. A stable HeLa cell line expressing His(6)-tagged SUMO-2 was established and used to label and purify novel endogenous SUMO-2 target proteins. Tagged forms of SUMO-2 were functional and localized predominantly in the nucleus. His(6)-tagged SUMO-2 conjugates were affinity-purified from nuclear fractions and identified by mass spectrometry. Eight novel potential SUMO-2 target proteins were identified by at least two peptides. Three of these proteins, SART1, heterogeneous nuclear ribonucleoprotein (RNP) M, and the U5 small nuclear RNP 200-kDa helicase, play a role in RNA metabolism. SART1 and heterogeneous nuclear RNP M were both shown to be genuine SUMO targets, confirming the validity of the approach.