色谱

色谱 ›› 2012, Vol. 30 ›› Issue (02): 170-177.DOI: 10.3724/SP.J.1123.2011.11020

• 研究论文 • 上一篇    下一篇

依赖色谱保留时间的智能化选择反应监测质谱方法的建立及其初步应用

毛心丽1,2, 卫军营2, 牛明2, 周廉淇2, 王雪颖1, 佟巍2, 秦伟捷2, 张养军2*, 钱小红2*   

  1. 1. 沈阳药科大学, 辽宁 沈阳 110015; 2. 蛋白质组学国家重点实验室, 北京蛋白质组研究中心, 军事医学科学院放射与辐射医学研究所, 北京 102206
  • 收稿日期:2011-11-11 修回日期:2011-12-05 出版日期:2012-02-28 发布日期:2012-03-22
  • 通讯作者: 钱小红,研究员,博士生导师,研究方向为蛋白质组学. Tel: (010)80727777-1231, E-mail: qianxh1@163.com. 张养军,博士,研究方向为蛋白质组新方法新技术. E-mail: zhangyangjun6314@yahoo.com.cn.
  • 基金资助:

    国家“863”计划项目(2009AA02Z205)、中国科学院重要方向项目(KSCX2-YW-G-050)、国家自然科学基金项目(81102345)和国家科技支撑计划项目(2009BADB9B02).

Establishment of retention time-dependent intelligent selected reaction monitoring mass spectrometric method and its application

MAO Xinli1,2, WEI Junying2, NIU Ming2, ZHOU Lianqi2, WANG Xueying1, TONG Wei2, QIN Weijie2, ZHANG Yangjun2*, QIAN Xiaohong2*   

  1. 1. Shenyang Pharmaceutical University, Shenyang 110015, China; 2. State Key Laboratory of Proteomics, Beijing Proteome Research Center, Beijing Institute of Radiation Medicine, Beijing 102206, China
  • Received:2011-11-11 Revised:2011-12-05 Online:2012-02-28 Published:2012-03-22

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321

被引次数

6

摘要： 建立了依赖色谱保留时间的智能化选择反应监测质谱方法,并与非依赖色谱保留时间的智能化选择反应监测质谱分析方法对不同体系(牛血清白蛋白酶切物、6种标准蛋白质混合物酶切物、腾冲嗜热菌蛋白提取液酶切物)的分析结果进行了系统比较。结果表明,引入色谱保留时间后的智能化选择反应监测质谱方法能够显著提高肽段及蛋白质的鉴定量,并且在复杂体系（如腾冲嗜热菌蛋白提取液酶切物）中效果尤为明显,鉴定到的肽段及蛋白质的覆盖率可分别达到目标肽段和蛋白质数量的89.62%和92.41%,并且灵敏度高、重复性好,能够实现对质荷比相同但保留时间有差异的肽段的准确鉴定。该方法将在复杂生物样本目标蛋白质组高通量、高灵敏度的鉴定、验证和确认中发挥独特作用。

关键词: 保留时间, 目标蛋白质组学, 腾冲嗜热菌, 液相色谱, 质谱, 智能化选择反应监测

Abstract: A method of liquid chromatographic retention time-dependent intelligent selected reaction monitoring mass spectrometry (iSRM MS) was established for the identification of targeted proteome, and was compared in detail with the method without liquid chromatographic retention time-dependent iSRM MS in the analysis results of different samples, such as the peptide mixtures from bovine serum albumin, six standard proteins or Thermoanaerobacter tengcongensis extract digested by trypsin. The results showed that the throughput of the identified peptides and the proteins was evidently increased with the method of liquid chromatographic retention time-dependent iSRM MS, especially in the complex sample such as Thermoanaerobacter tengcongensis extract. The coverage of the identified peptides and proteins from Thermoanaerobacter tengcongensis extract was reached 89.62% and 92.41% of the number of targeted peptides and proteins respectively. Higher sensitivity and reproducibility were also obtained. In addition, peptides with the same m/z but different retention times could be identified accurately when the method was used. With the higher throughput, better sensitivity and excellent reproducibility, the method of liquid chromatographic retention time-dependent iSRM MS can play an important role in the identification of targeted proteomes of complex biological samples in the future.

Key words: intelligent selected reaction monitoring (iSRM), mass spectrometry (MS), retention time, targeted proteomics, Thermoanaerobacter tengcongensis, liquid chromatography (LC)