基质辅助激光解析电离-串联飞行时间质谱与电喷雾离子化-四极杆-飞行时间质谱分析蛋白质  

Comparison of Performance of Matrix-Assisted Laser Desorption/Ionization-Time of Flight-Time of Flight and Electrospray Ionization-Quadrupole-Time of Flight for Qualification and Quantification of Proteins

作　　者：倪国新[1] 朱镭 马小土[1] 徐学敏[1,2] 胡晓芳[2] 刘建华[2] 李伟 

机构地区：[1]上海交通大学系统生物医学研究院教育部系统生物医学重点实验室,上海200240 [2]上海交通大学生命科学技术学院,上海200240

出　　处：《分析化学》2009年第9期1291-1296,共6页Chinese Journal of Analytical Chemistry

基　　金：国家重点基础研究发展规划项目(No.2006CB0D0100)资助

摘　　要：为比较基质辅助激光解析电离-串联飞行时间质谱(MALDI-TOF-TOF)和电喷雾离子化-四极杆-飞行时间质谱(ESI-Q-TOF)在蛋白质定性和定量分析方面的性能,分别利用两种仪器对用于相对和绝对定量的等量异位标签(iTRAQ)标记的雌激素刺激前后MCF7细胞内的蛋白质水解产生的多肽进行了定性和定量分析。结果表明,用MALDI-TOF-TOF和ESI-Q-TOF方法分别鉴定出1086种和848种蛋白质,其中相同的蛋白质为633种;MALDI-TOF-TOF和ESI-Q-TOF方法分别找到38种和33种表达差异在0.5倍以上的蛋白质,其中3种为相同的蛋白质。实验数据说明,两种仪器在蛋白质定性和定量分析方面有很大的互补性,将两种仪器结合使用,不仅能够显著提高蛋白质定性和定量分析的覆盖率,而且可提高分析的置信度。To compare the performance of MALDI-TOF-TOF and ESI-Q-TOF in protein qualification and quantification,isobaric tags for relative and absolute quantification（iTRAQ） labeled peptides derived from trypsin-digested proteins in MCF7 cells with or without estrogen treatment were identified and quantified using MALDI-TOF-TOF and ESI-Q-TOF respectively.The experimental result showed that 1086 and 848 proteins were identified by MALDI-TOF-TOF and ESI-Q-TOF respectively,wherein 633 proteins were homolog;38 and 33 proteins were found to be differentially expressed by more than 0.5 fold by MALDI-TOF-TOF and ESI-Q-TOF respectively,wherein 3 proteins were the same proteins.The experimental data indicate that these two kinds of instruments were complementary both in peptide identification and relative quantification.Paralleled use of MALDI-TOF-TOF and ESI-Q-TOF significantly increased both the coverage and the confidence level of identified and quantified proteins.

关 键 词：等量异位标签 基质辅助激光解析电离-串联飞行时间质谱 电喷雾离子化-四极杆-飞行时间质谱 定性分析 定量分析 

分 类 号：O657.63[理学—分析化学] TQ936.1[理学—化学]