Label-free detection of differential protein expression by LC/MALDI mass spectrometry
Label-free detection of differential protein expression by LC/MALDI mass spectrometry
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DOI：
10.1021/pr700705u
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发表时间：
2008-06-01
影响因子：
4.4
通讯作者：
James, Ian T.
中科院分区：
文献类型：
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作者：
Neubert, Hendrik;Bonnert, Timothy P.;James, Ian T.
蛋白质丰度在疾病或实验扰动期间的变化越来越多地被无标记的LC/MS方法分析。在这里，我们使用LC/MALDI MS对以阿拉伯糖、果糖或葡萄糖为碳源的大肠杆菌培养物进行无标记蛋白质表达差异检测。说明了MALDI的优点，如只检测单电荷离子，以及MALDI板存档，便于回溯性的MS/MS数据收集。用Rosetta分析系统对大肠杆菌裂解物的胰酶消化反相层析的MALDI光谱进行了比对和定量。在跨越3个数量级的信号强度的所有LC/MALDI运行中检测到大约5000个肽信号。在所有技术复制中，所有信号在整个强度范围内的平均变异系数被发现为
Protein abundance changes during disease or experimental perturbation are increasingly analyzed by label-free LC/MS approaches. Here we demonstrate the use of LC/MALDI MS for label-free detection of protein expression differences using Escherichia coli cultures grown on arabinose, fructose or glucose as a carbon source. The advantages of MALDI, such as detection of only singly charged ions, and MALDI plate archiving to facilitate retrospective MS/MS data collection are illustrated. MALDI spectra from RP chromatography of tryptic digests of the E. coli lysates were aligned and quantitated using the Rosetta Elucidator system. Approximately 5000 peptide signals were detected in all LC/MALDI runs spanning over 3 orders of magnitude of signal intensity. The average coefficients of variation for all signals across the entire intensity range in all technical replicates were found to be