论文部分内容阅读
质谱法鉴定双向电泳胶内蛋白质是目前蛋白质组研究中最重要的分析方法．采用电喷雾离子阱质谱法对人肝癌细胞系ＢＥＬ－７４０４双向电泳后的胶内蛋白质进行分析鉴定．蛋白质经双向电泳分离，从凝胶上切割后，经过胰蛋白酶胶内酶解，通过液相色谱－电喷雾离子阱质谱分析肽谱和各肽段的氨基酸序外．根据肽谱和序列信息，经数据库检索，结合双向电泳分离获得的蛋白质分子量和等电点信息，对９种胶内蛋白质进行了鉴定．获得的氨基酸覆盖率为对２１％～７２．２％．其中两种蛋白质在ＳＷＩＳＳ－２ＤＰＡＧＥ的肝细胞蛋白质组数据库中尚未报道．在此基础上，进一步探讨了所鉴定的蛋白质与肝癌发生的关系．这一方法的应用，为与疾病相关的蛋白质组研究提供了有效和快速的分析鉴定方法．
Mass spectrometry identification of proteins in two-dimensional electrophoresis gels is the most important analytical method in proteome studies. Electrophoretic ion trap mass spectrometry was used to analyze and identify the in-gel proteins after two-dimensional electrophoresis of human hepatocellular carcinoma cell line BEL-7404. The protein was separated by two-dimensional electrophoresis. After being cut from the gel, it was enzymatically digested with trypsin and analyzed by liquid chromatography-electrospray ion trap mass spectrometry to analyze the peptide profile and amino acid sequence of each peptide. Based on the peptide spectra and sequence information, the database was searched and the molecular weight and isoelectric point information of the protein obtained by two-dimensional electrophoresis separation were used to identify nine proteins in the gel. The coverage of amino acids obtained is between 21% and 72.2%. Two of these proteins have not been reported in the SWISS-2DPAGE hepatocyte proteome database. On this basis, the relationship between the identified proteins and the occurrence of liver cancer was further explored. The application of this method provides an effective and rapid analysis and identification method for disease-related proteome research.