受精15.5天胎鼠血清和新生鼠血清蛋白质组学比较研究

发布时间：2018-03-07 20:32

本文选题：血浆蛋白质组　切入点：发育　出处：《北京协和医学院》2011年博士论文　论文类型：学位论文

【摘要】：血液是人体的重要体液,包含了来自全身各组织分泌的蛋白和细胞死亡渗出的蛋白,在某种程度上可以说血液是人体的窗口。在胎儿发育过程中,尤其是胎儿发育中后期,各器官无论在形态还是在功能都发生了巨大的变化。基于液质联用的蛋白质组学分析技术能对蛋白质——生命功能的具体执行者,进行高通量、大规模的全面研究。通过使用液质联用技术对不同时期胎鼠血清进行蛋白质组学鉴定分析,能够更好的了解生命发育的过程。考虑到胎鼠血清个体差异,分别对受精15.5天胎鼠和新生鼠血清的个体样本和混合样本进行质谱鉴定。以混合样本三次质谱鉴定的变异系数(作为实验方法的差异)为对照,计算个体样本变异系数与混合样本变异系数的比值,获得每个蛋白在去除质谱仪器等实验误差造成的影响后,在蛋白质组学水平上进行个体差异性评价。应用毛细管液相色谱(反相)和质谱联用方法,在质谱鉴定可信度为99%的条件下,在个体样本中,共鉴定的669个蛋白,对鉴定的蛋白进行双侧T检验,在95%的可信区间内分别有35个蛋白显著性上升和58个蛋白显著性降低。对鉴定的蛋白进行功能注释,并对在血浆中参与运输、防御、酶活性调节、凝血等生物学过程的蛋白进行分析,共发现9种补体分子,均上升,其中6种出现显著性上升；酶活性调节物分子中4种显著性下降,13种显著性上升；载脂蛋白分子显著减少,11种载脂蛋白分子中8种显著性下降,1种显著性升高；同时发现不同种类血红蛋白分子出现改变。蛋白翻译后修饰对蛋白结构和功能起着重要的调节作用。通过对不同时期血清混合样本蛋白的翻译后修饰的研究,发现了50种不同的修饰。通过对修饰位点的定量比较,发现受精15.5天和新生鼠血清蛋白翻译后修饰的状态不同,为以后研究翻译后修饰在血清发育中的作用提供线索。
[Abstract]:Blood is the important body fluid of the human body. It contains proteins secreted from various tissues of the whole body and proteins exudated by cell death. To some extent, blood is the window of the human body. In the course of fetal development, especially in the middle and late stages of fetal development, Great changes have taken place in both morphology and function of the organs. Proteomics analysis based on liquid-mass spectrometry can perform high throughput on the specific implementers of protein-life function. Through the proteomics identification and analysis of fetal rat serum at different stages, we can better understand the process of life development. Taking into account the individual difference of fetal rat serum, the individual and mixed samples of fetal mouse and newborn mouse serum on the 15.5 day of fertilization were identified by mass spectrometry, and the coefficient of variation (as the difference of experimental method) of the three times mass spectrometry identification of mixed sample was compared. The ratio of the coefficient of variation of individual samples to that of mixed samples was calculated and the individual differences were evaluated at proteomics level after removing the effects of experimental errors such as mass spectrometer. Capillary liquid chromatography (RP-MS) and mass spectrometry were used to detect 669 proteins in individual samples under the condition that the reliability of mass spectrometry was 99%. 35 proteins increased significantly and 58 proteins decreased significantly in the confidence interval of 95%. The identified proteins were annotated and regulated to participate in transport, defense and enzyme activity in plasma. In the analysis of proteins in biological processes such as coagulation, 9 complement molecules were found to be increased, among which 6 were significantly increased, 4 of the enzyme activity regulators were significantly decreased and 13 were significantly increased. Apolipoprotein molecules decreased significantly in 8 out of 11 apolipoprotein molecules and increased significantly in 1 out of 11 apolipoprotein molecules, and different hemoglobin molecules were found to change. Posttranslational modification of proteins plays an important role in regulating the structure and function of proteins. Through the study of post-translational modification of serum mixed sample proteins at different stages, 50 different modifications were found. It was found that the posttranslational modification of serum proteins in 15.5 days of fertilization was different from that in newborn mice, which provided clues for further study on the role of posttranslational modification in serum development.
【学位授予单位】：北京协和医学院
【学位级别】：博士
【学位授予年份】：2011
【分类号】：R321.2;Q51

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