Small ubiquitin-like modifier 3-modified proteome regulated by brain ischemia in novel small ubiquitin-like modifier transgenic mice: putative protective proteins/pathways.
Small ubiquitin-like modifier 3-modified proteome regulated by brain ischemia in novel small ubiquitin-like modifier transgenic mice: putative protective proteins/pathways.
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DOI：
10.1161/strokeaha.113.004315
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发表时间：
2014-04
期刊：
影响因子：
8.3
通讯作者：
Paschen W
中科院分区：
文献类型：
--
作者：
Yang W;Sheng H;Thompson JW;Zhao S;Wang L;Miao P;Liu X;Moseley MA;Paschen W
SUMO缀合是与许多人类疾病相关的翻译后修饰。SUMO修饰的蛋白质组的表征对于确定SUMO缀合与此类疾病之间的机制联系至关重要。这对于SUMO 2/3缀合尤其明显，SUMO 2/3缀合在脑缺血/中风后被大量激活，并且被认为是一种保护性反应。本研究的目的是使用一种新的SUMO转基因小鼠对脑缺血调控的SUMO 3修饰蛋白质组进行全面分析。为了能够在体内进行SUMO蛋白质组学分析，我们产生了条件性表达标记的SUMO 1 -3旁系同源物的转基因小鼠。转基因小鼠前脑缺血10分钟，再灌注1小时。通过抗FLAG亲和纯化富集SUMO 3缀合的蛋白，并通过LC-MS/MS分析。SUMO转基因小鼠的表征表明，所有3个标记的SUMO旁系同源物都具有功能活性，并且外源性SUMO的表达不改变内源性SUMO化机制。蛋白质组学分析确定了112个假定的SUMO 3底物，其中91个候选物在缺血组中比假手术组更丰富。数据分析揭示了具有假定神经保护功能的过程/途径，包括糖皮质激素受体信号传导、RNA加工和SUMO化依赖的泛素缀合。通过SUMO化调节的已鉴定蛋白/通路可能是理解SUMO化与神经保护相关机制的关键，从而为治疗干预提供新的有希望的靶点。这种新的转基因小鼠将成为分析人类疾病模型中SUMO修饰蛋白质组的宝贵平台，从而有助于将SUMO化与病理过程联系起来。
SUMO conjugation is a post-translational modification associated with many human diseases. Characterization of the SUMO-modified proteome is pivotal to defining the mechanistic link between SUMO conjugation and such diseases. This is particularly evident for SUMO2/3 conjugation, which is massively activated after brain ischemia/stroke, and is believed to be a protective response. The purpose of this study was to perform a comprehensive analysis of the SUMO3-modified proteome regulated by brain ischemia using a novel SUMO transgenic mouse. To enable SUMO proteomics analysis in vivo, we generated transgenic mice conditionally expressing tagged SUMO1-3 paralogues. Transgenic mice were subjected to 10 minutes forebrain ischemia and 1 hour of reperfusion. SUMO3-conjugated proteins were enriched by anti-FLAG affinity purification and analyzed by LC-MS/MS. Characterization of SUMO transgenic mice demonstrated that all 3 tagged SUMO paralogues were functionally active, and expression of exogenous SUMOs did not modify the endogenous SUMOylation machinery. Proteomics analysis identified 112 putative SUMO3 substrates of which 91 candidates were more abundant in the ischemia group than the sham group. Data analysis revealed processes/pathways with putative neuroprotective functions, including glucocorticoid receptor signaling, RNA processing and SUMOylation-dependent ubiquitin conjugation. The identified proteins/pathways modulated by SUMOylation could be the key to understanding the mechanisms linking SUMOylation to neuroprotection, and thus provide new promising targets for therapeutic interventions. The new transgenic mouse will be an invaluable platform for analyzing the SUMO-modified proteome in models of human disorders, and thereby help to mechanistically link SUMOylation to the pathological processes.