Distinct and overlapping sets of SUMO-1 and SUMO-2 target proteins revealed by quantitative proteomics
Distinct and overlapping sets of SUMO-1 and SUMO-2 target proteins revealed by quantitative proteomics
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DOI：
10.1074/mcp.m600212-mcp200
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发表时间：
2006-12-01
影响因子：
7
通讯作者：
Lamond, Angus I.
中科院分区：
文献类型：
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作者：
Vertegaal, Alfred C. O.;Andersen, Jens S.;Lamond, Angus I.
脊椎动物中的小泛素样修饰物（SUMO）家族包括三个不同的家族成员，其作为翻译后修饰缀合至靶蛋白。SUMO-2和-3几乎相同，但与SUMO-1有很大不同。我们使用定量蛋白质组学研究SUMO-1和SUMO-2的靶蛋白偏好。建立稳定表达His 6-SUMO-1或His 6-SUMO-2的HeLa细胞。这些细胞系和对照HeLa细胞用稳定的精氨酸同位素标记，并使用固定化金属亲和层析从裂解物中富集His 6-SUMO。共鉴定了53个SUMO结合蛋白，其中包括44个新的SUMO靶点。25种蛋白质优先与SUMO-1缀合，19种蛋白质优先与SUMO-2缀合，9种蛋白质与SUMO-1和SUMO-2两者缀合。通过免疫印迹证实SART 1具有相似水平的SUMO-1-和SUMO-2-连接形式。因此，SUMO-1和SUMO-2显示具有不同的和重叠的靶蛋白组，表明SUMO-1和SUMO-2可能具有冗余和非冗余的细胞功能。有趣的是，25个SUMO-1结合蛋白中有14个含有锌指。虽然这两个SUMO家族成员在许多细胞过程中发挥作用，我们的数据表明，SUMO化与转录密切相关，因为近三分之一的确定的靶蛋白是公认的转录调节因子。
The small ubiquitin-like modifier ( SUMO) family in vertebrates includes three different family members that are conjugated as post-translational modifications to target proteins. SUMO-2 and -3 are nearly identical but differ substantially from SUMO-1. We used quantitative proteomics to investigate the target protein preferences of SUMO-1 and SUMO-2. HeLa cells were established that stably express His 6-SUMO-1 or His 6-SUMO-2. These cell lines and control HeLa cells were labeled with stable arginine isotopes, and His 6-SUMOs were enriched from lysates using immobilized metal affinity chromatography. 53 SUMO-conjugated proteins were identified, including 44 novel SUMO targets. 25 proteins were preferentially conjugated to SUMO-1, 19 were preferentially conjugated to SUMO-2, and nine proteins were conjugated to both SUMO-1 and SUMO-2. SART1 was confirmed by immuno-blotting to have both SUMO-1- and SUMO-2-linked forms at similar levels. SUMO-1 and SUMO-2 are thus shown to have distinct and overlapping sets of target proteins, indicating that SUMO-1 and SUMO-2 may have both redundant and non-redundant cellular functions. Interestingly, 14 of the 25 SUMO-1-conjugated proteins contain zinc fingers. Although both SUMO family members play roles in many cellular processes, our data show that sumoylation is strongly associated with transcription because nearly one-third of the identified target proteins are putative transcriptional regulators.