摘要
随着“一小时酵母蛋白质组”的实现，短梯度下实现蛋白质组深度覆盖成为可能．利用全新Q-0T-qlT三合一质谱系统进行“一小时蛋白质组”分析与优化．50min有效梯反，单次实验分别从1μg和50 ng HeLa全蛋白中鉴定到20860和14100条肽段，对应到3865和2877个非冗余蛋白，而目前的文献报道至少需要2～3h．同时，考察了Q-OT-qlT碎裂模式、检测方法、最大注入时间和自动增益控制等参数对蛋白质组快速分析的影响，证明了不同采集方法间的互补性，阐述了不同扫描参数对鉴定结果的影响．此外，还讨论了Q-OT-qIT的并列运行原理和扫描组合模式，为不同实验目的和样本类型的蛋白质组快速分析奠定了基础．
With the realization of ＂the one hour yeast proteome＂, in-deepth proteome under rapid LC-MS/MS gradient is achieved. The current study utilized a novel Q-OT-qlT mass spectrometer to analyze and optimize ＂the one hour proteome＂. Within 50 vain LC-MS/MS gradient, a total of 20860 and 14100 unique peptides were identified from 1 μg and 50 ng HeLa digest, corresponding to 3865 and 2877 proteins, respectively. Similar result from current reports used at least 2-3 h gradient. The effects of various collision modes, detectors, maximum injection times and automatic gain control settings were further evaluated, and the complementarity between different acquisition methods and scan parameters were described. Moreover, the massive parallelization of Q-OT-qlT was discussed, providing useful information for rapid proteome analysis.
出处
《中国科学：生命科学》
CSCD
北大核心
2014年第5期528-536,共9页
Scientia Sinica(Vitae)
