基于不同裂解液的蛋白质组样品制备方法的比较研究  

Comparative Study of Three Methods of Sample Preparation for Proteomics Research

作　　者：孙宁宁[1] 王子健[1] 高祥[1] 程欣[1] 孙万春[2] 杨静波[1] 刘宁[1] 

机构地区：[1]吉林大学第二医院研究中心,吉林省重大疾病分子与化学遗传学重点实验室,长春130041 [2]吉林大学人兽共患病研究所,人兽共患病研究教育部重点实验室,长春130062

出　　处：《分析化学》2015年第6期808-813,共6页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金(Nos.21175055,81472030);吉林省科技发展计划(Nos.20110739,20150204001YY);吉林大学白求恩计划B(No.2012210)项目的支持~~

摘　　要：蛋白质组学研究所面临的首要问题就是蛋白质组的样品制备,而不同的样品制备方案针对细胞、组织等样品的裂解能力,以及对不同种类蛋白的溶解能力会有很大的差异。本研究以培养的293T细胞为对象,利用质谱分析鉴定蛋白的方法,比较了采用3种常用的蛋白质组样品制备方法(Triton X-100法、尿素法和TRIzol法)所提取蛋白种类的差异,并利用生物信息学工具对所得数据进行了分析。结果表明,Triton X-100法和尿素法所提取蛋白的鉴定数目接近,而TRIzol法所提取蛋白的鉴定数目比二者减少约8%。从所鉴定的蛋白种类来看,3种方法相差较大,仅约有32%的蛋白为3种方法所共有,生物学功能也不尽相同。本研究为评价各种蛋白质组样品制备方法提供了一种快速、有效、全面的研究方法。Protein sample preparation is a vital issue for proteomics research. Due to different physical and chemical properties of reagents used for sample preparation, the abilities of various reagents to disrupt cell or tissue, as well as to solubilize a variety of proteins, are very different. In the present study, three sample preparation methods widely used in proteomics researches （Triton X-100 method, urea method and TRIzol method） were compared by analyzing proteins isolated from cultured 293T cells using mass spectrometric technique, followed by bioinformatics analyses. The results indicated that the number of identified proteins extracted by Triton X-100 method was almost the same as that by urea method, whereas the number of identified proteins extracted by TRIzol m^thod was approximately 8 percent smaller than those by either Triton X-100 or urea method. A large difference was found among the protein categories identified by three extraction methods with approximate only 32 percent of total proteins identified by all the three methods. The profiles of proteins prepared by three methods were compared and further analyzed using functional classification software. This study provides a rapid, effective and comprehensive tool for evaluating the sample preparation methods for proteomies study.

关 键 词：蛋白质组 样品制备 尿素 曲拉通X-100 TRIZOL 液相色谱-质谱联用 

分 类 号：Q51[生物学—生物化学]