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目的构建小鼠肝脏磷酸化蛋白质组并对磷酸化激酶进行分析,为完善小鼠激酶磷酸化调控网络提供有价值的信息。方法对正常小鼠肝组织总蛋白提取液进行FASP酶切,用Ti O2富集磷酸化肽段,为降低样本的复杂度,对富集到的磷酸化肽段进行反相色谱分离后,质谱鉴定样本中的磷酸化蛋白质组,对鉴定到的磷酸化修饰的激酶进行分析,提供新鉴定到磷酸化修饰位点的信息。结果与结论成功构建了高效的鉴定小鼠肝磷酸化蛋白质组的方法,共鉴定到1533个磷酸化蛋白质,从中确认5386个磷酸化位点和4553个磷酸化肽段,其中包含116磷酸化修饰的激酶,并于发生磷酸化修饰的激酶中成功鉴定到126个新的磷酸化修饰位点,为完善小鼠肝磷酸化信号调控网络提供了有价值的信息。
Objective To construct mouse liver phosphoproteome and to analyze phosphorylated kinase, so as to provide valuable information for improving mouse phosphorylation regulatory network. Methods The normal mouse liver total protein extract was subjected to FASP digestion, and the phosphorylated peptides were enriched with Ti O2. To reduce the complexity of the samples, the phosphorylated peptides were separated by reversed-phase chromatography. Mass spectra Identify the phosphorylated proteome in the sample and analyze the identified phosphorylated kinase to provide information for newly identified phosphorylation sites. RESULTS AND CONCLUSION: An efficient method for identification of mouse hepatic phosphoproteome was successfully constructed. A total of 1533 phosphorylated proteins were identified, among which 5386 phosphorylation sites and 4553 phosphorylated peptides were identified, including 116 phosphorylation modification , And successfully identified 126 new phosphorylation sites in phosphorylation-modified kinases, which provided valuable information for improving the hepatic phosphorylation regulatory network in mice.