Sumo-dependent substrate targeting of the SUMO protease Ulp1.
Sumo-dependent substrate targeting of the SUMO protease Ulp1.
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DOI：
10.1186/1741-7007-9-74
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发表时间：
2011-10-28
期刊：
影响因子：
5.4
通讯作者：
Kerscher O
中科院分区：
文献类型：
--
作者：
Elmore ZC;Donaher M;Matson BC;Murphy H;Westerbeck JW;Kerscher O
在酿酒酵母中，必需的小泛素样修饰物（SUMO）蛋白酶Ulp 1负责从特定靶蛋白中去除SUMO/Smt 3，并将前体SUMO加工成具有缀合能力的形式。Ulp 1主要定位于核孔复合物，但也已被证明在分裂细胞的芽颈处解偶联sumoylated septins。Ulp 1是如何定向于芽颈定位的septins和其他细胞质去缀合靶标的尚不清楚。使用结构/功能的方法，我们着手阐明底物靶向所需的Ulp 1的功能。为了帮助我们的研究，我们利用了Ulp 1的催化失活突变体，该突变体在分裂酵母细胞的隔蛋白环处大大富集。我们发现，本地化的Ulp 1的septins需要SUMO和特定的结构特征的Ulp 1的催化结构域。我们的分析确定了一个218个氨基酸的底物捕获突变体Ulp 1（3）（C580 S）的催化结构域，这是必要的和足够的septin定位。我们还利用Ulp 1（3）（C580 S）的靶向和SUMO结合特性从细胞提取物中纯化Smt 3修饰的蛋白质。我们的研究提供了新的见解如何Ulp 1 SUMO蛋白酶是积极针对其底物在体内和体外。此外，我们发现底物捕获Ulp 1（3）（C580 S）与人SUMO 1，SUMO 2和SUMO 2链强烈相互作用，使其成为分析和纯化SUMO修饰蛋白质的潜在有用工具。
In the yeast Saccharomyces cerevisiae, the essential small ubiquitin-like modifier (SUMO) protease Ulp1 is responsible for both removing SUMO/Smt3 from specific target proteins and for processing precursor SUMO into its conjugation-competent form. Ulp1 localizes predominantly to nuclear pore complexes but has also been shown to deconjugate sumoylated septins at the bud-neck of dividing cells. How Ulp1 is directed to bud-neck localized septins and other cytoplasmic deconjugation targets is not well understood. Using a structure/function approach, we set out to elucidate features of Ulp1 that are required for substrate targeting. To aid our studies, we took advantage of a catalytically inactive mutant of Ulp1 that is greatly enriched at the septin ring of dividing yeast cells. We found that the localization of Ulp1 to the septins requires both SUMO and specific structural features of Ulp1's catalytic domain. Our analysis identified a 218-amino acid, substrate-trapping mutant of the catalytic domain of Ulp1, Ulp1(3)(C580S), that is necessary and sufficient for septin localization. We also used the targeting and SUMO-binding properties of Ulp1(3)(C580S) to purify Smt3-modified proteins from cell extracts. Our study provides novel insights into how the Ulp1 SUMO protease is actively targeted to its substrates in vivo and in vitro. Furthermore, we found that a substrate-trapping Ulp1(3)(C580S) interacts robustly with human SUMO1, SUMO2 and SUMO2 chains, making it a potentially useful tool for the analysis and purification of SUMO-modified proteins.
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DOI：
10.1083/jcb.200212052
发表时间：
2003-03-31
期刊：
The Journal of cell biology
影响因子：
--
作者：
Li SJ;Hochstrasser M
通讯作者：
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影响因子：
7.8
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影响因子：
64.5
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影响因子：
7.8
作者：
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Hochstrasser, Mark
DOI：
10.1074/mcp.m700173-mcp200
发表时间：
2008-01
期刊：
Molecular & cellular proteomics : MCP
影响因子：
--
作者：
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