The one hour yeast proteome.
The one hour yeast proteome.
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DOI：
10.1074/mcp.m113.034769
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发表时间：
2014-01
期刊：
影响因子：
--
通讯作者：
Coon JJ
中科院分区：
文献类型：
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作者：
Hebert AS;Richards AL;Bailey DJ;Ulbrich A;Coughlin EE;Westphall MS;Coon JJ
我们描述了在短短一个多小时的优化分析中对酵母蛋白质组的全面分析。我们实现了这种加快蛋白质组表征与改进的样品制备，色谱分离，并通过使用新的Orbitrap混合质谱仪配备了质量过滤器，碰撞池，高场Orbitrap分析仪，最后，双细胞线性离子阱分析仪（Q-OT-qIT，Orbitrap融合）。该系统提供高达20 Hz的MS 2采集速度，并在一秒钟内检测多达19个肽序列。在1.3小时的色谱方法中，Q-OT-qIT杂合体收集了平均13，447次MS 1和80，460次MS 2扫描（每次运行），以产生43，400（x 100）个肽光谱匹配和34，255（x 100）个具有独特氨基酸序列的肽（1%错误发现率（FDR））。平均而言，每一个小时的分析实现了3，977个蛋白质的检测（1% FDR）。我们的结论是，质谱仪扫描速率的进一步改进可以在几个小时内对人类蛋白质组进行全面分析。
We describe the comprehensive analysis of the yeast proteome in just over one hour of optimized analysis. We achieve this expedited proteome characterization with improved sample preparation, chromatographic separations, and by using a new Orbitrap hybrid mass spectrometer equipped with a mass filter, a collision cell, a high-field Orbitrap analyzer, and, finally, a dual cell linear ion trap analyzer (Q-OT-qIT, Orbitrap Fusion). This system offers high MS2 acquisition speed of 20 Hz and detects up to 19 peptide sequences within a single second of operation. Over a 1.3 h chromatographic method, the Q-OT-qIT hybrid collected an average of 13,447 MS1 and 80,460 MS2 scans (per run) to produce 43,400 (x̄) peptide spectral matches and 34,255 (x̄) peptides with unique amino acid sequences (1% false discovery rate (FDR)). On average, each one hour analysis achieved detection of 3,977 proteins (1% FDR). We conclude that further improvements in mass spectrometer scan rate could render comprehensive analysis of the human proteome within a few hours.