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目的 寻找并验证Ubr1泛素化底物蛋白大规模筛选方法的可行性.方法 通过对啤酒酵母RJD347(Ubr1野生型)和AVY26(Ubr1缺失型)的差异蛋白质组学比较,分析酵母中可能存在的Ubr1直接或间接作用的底物蛋白.进一步通过外源表达实验验证差异蛋白与Ubr1之间的相关性.结果 组学研究得到249个差异表达蛋白,其中145个蛋白在AVY26(Ubr1缺失型)中表达上调.选取其中40个并外源表达验证5个差异蛋白MLC2、SCD6、ARG1、HOG1及RTF1与Ubr1具有相关性,受泛素连接酶Ubr1的泛素化调控.结论 建立Ubr1泛素化底物候选蛋白库,同时验证出5个潜在Ubr1泛素化底物蛋白.“,”Objective To justify the rationality of proteomic screening on a large scale and identify potential substrates of Ubr1 with the approach.Methods In this work,comparative proteomic analysis between RJD347 (Ubr1 wild type) and AVY26 (Ubr1 deletion type) has been performed to screen the possible substrate proteins of Ubrl.Moreover,the correlation between the differentially expressed proteins and Ubr1 was further verified by protein work.Results In total,249 proteins which were differentially expressed were identified,of which 145 proteins were up-regulated in AVY26.Furthermore,the protein work confirmed 5 proteins including MLC2,SCD6,ARG1,HOG1 and RTF1 that were closely related to Ubr1.Conclusions A massive database of 249 proteins that are differentially expressed between RJD347 and AVY26 is established.Five potential substrates of Ubr1 ubiquitination proteins is identified,which provides the experimental basis for further understanding of biological processes of Ubr1.