大规模提高泛素化肽段鉴定数量的新方法研究

项目名称： 大规模提高泛素化肽段鉴定数量的新方法研究

项目编号： No.31200612

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 生物物理、生化与生物分子学、生物力学与组织工程

项目作者： 肖传乐

作者单位： 暨南大学

项目金额： 20万元

中文摘要： 泛素化在蛋白质降解中起着重要的作用。然而，蛋白质泛素化修饰存在着普遍性和动态性，它使得大规模的检测泛素化修饰非常困难。以致寻求有效的方法确定细胞和组织内泛素化谱是当今功能蛋白质组学研究的重要内容。目前鉴定泛素化肽段数量还比较低，因此需要一种新的方法能够提高其数量。本项目是生物信息学理论与生物质谱实验的有机结合，试图通过LC离线分离肽段和泛素化抗体富集泛素化肽段，并对富集后的各组分样品等分成多个组分进行多次质谱分析：第一次质谱实验为普通质谱试验，通过计算生物学方法分析第一次质谱数据，预测第二次质谱分析中每个时间点应该监测的母离子峰列表，通过该动态列表控制第二次质谱实验，依次类推，重复多次计算分析和实验，从而探索出大规模提高泛素化肽段鉴定数量和定量数量的新方法。该方法可以有效利用生物质谱的灵敏度，鉴定到普通质谱分析没有检测到质谱信息，为翻译后修饰提供方法参考。

中文关键词： 泛素化；蛋白质质谱鉴定方法；基于结果驱动质谱方法；母离子峰识；

英文摘要： Ubiquitination plays an important role in protein degradation . Since ubiquitin modification of protein is the character of universal and dynamic, it makes large scale identification of ubiquitin peptide very difficult，so some research which focus on an effective method for largely detection ubiquitin modification is important content in function proteomics research. This project organically combine bioinformatics theory with mass spectrometry(MS) experiment and attempts to separate peptides by offline LC and enrich ubiquitin peptides by ubiquitin antibody. each enriched sample is evenly divided into several components to supply some times MS experiment: the first MS experiment is ordinary MS analysis , peptide ion peak list with MS analysis time which should be detected in the second parallel MS experiment is predicted from the first MS data by computational biology and control the second MS experiment . Peptide ion list of the third MS experiment is gained from two times MS data. Thereby a new method with largly increasing idenfication number of ubiquitin peptides is studied by the repeat above procedure. This method can effectively use the sensitivity of MS, detect large information whose information is not detect MS information in the ordinary MS analysis and provide new methods for posttranslational modifi

英文关键词： Ubiquitination；MS identification algorithms of protein；MS strategy based on result-driven；the identification of the parent ion spectrum；