New algorithm for 15N/14N quantitation with LC-ESI-MS using an LTQ-FT mass spectrometer
New algorithm for 15N/14N quantitation with LC-ESI-MS using an LTQ-FT mass spectrometer
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DOI：
10.1021/pr060105m
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发表时间：
2006-08-04
影响因子：
4.4
通讯作者：
Karger, Barry L.
中科院分区：
文献类型：
--
作者：
Andreev, Victor P.;Li, Lingyun;Karger, Barry L.
描述了一种用于复杂蛋白质组样品中相对蛋白质丰度 N-15/N-14 定量的新算法 (QN)。 QN 利用混合质谱仪 LTQ-FT MS 的高分辨率、质量精度和通量。肽定量基于 MS 峰强度（在 FT MS 中测量），而肽鉴定则在 MS/MS 模式下进行（在 LTQ 线性离子阱中测量）。新颖的评分程序提高了蛋白质丰度的准确性，从而可以过滤不太可靠的肽丰度测量值。 QN 的性能通过对以一氧化碳和甲醇为底物生长的 M. acetivorans C2A 培养物的相对定量分析来说明。对大约 1000 种蛋白质进行了定量，蛋白质丰度比的平均 CV 为 9%。 QN 无需人工干预即可进行定量，不需要高处理能力，并生成与蛋白质组数据发布指南兼容的文件。
A new algorithm (QN) for the N-15/N-14 quantitation of relative protein abundances in complex proteomic samples is described. QN takes advantage of the high resolution, mass accuracy and throughput of the hybrid mass spectrometer LTQ-FT MS. Peptide quantitation is based on MS peak intensity (measured in the FT MS), while peptide identification is performed in the MS/MS mode (measured in the LTQ linear ion trap). Accuracy of the protein abundance is enhanced by a novel scoring procedure, allowing filtering of less reliable measurements of peptide abundances. The performance of QN is illustrated in the relative quantitative analysis of M. acetivorans C2A cultures grown with carbon monoxide vs methanol as substrate. Roughly 1000 proteins were quantitated with an average CV of 9% for the protein abundance ratios. QN performs quantitation without manual intervention, does not require high processing power, and generates files compatible with the Guidelines for Proteomic Data Publication.