Assessing reproducibility of a protein dynamics study using in vivo labeling and liquid chromatography tandem mass spectrometry
Assessing reproducibility of a protein dynamics study using in vivo labeling and liquid chromatography tandem mass spectrometry
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DOI：
10.1021/ac048204b
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发表时间：
2005-05-01
影响因子：
7.4
通讯作者：
Pandey, A
中科院分区：
文献类型：
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作者：
Molina, H;Parmigiani, G;Pandey, A
测量细胞中蛋白质丰度响应于诸如生长因子或药物的刺激的动态需要分析多于一个时间点。蛋白质组学方法传统上用于一次仅测量一种状态，因为定量是困难的，特别是当质谱法用作读数时。同位素标记的试剂最近已经被引入，其允许通过质谱法比较两种或三种不同的状态。在这里，我们评估了一个实验，同时测量三个状态，通过稳定同位素标记的细胞与氨基酸在细胞培养（SILAC）使用轻，中，重版本的氨基酸。本研究的主要目的是评估此类实验与液相色谱串联质谱（LC-MS/MS）结合的重现性。我们的研究结果表明，它是可能的，以获得可重复的定量数据来研究蛋白质动力学的基础上，我们的分析超过220肽集来自。来自3次不同LC-MS/MS运行的20种蛋白质。
Measuring dynamics of proteins abundance in cells in response to stimuli such as growth factors or drugs requires analysis of more than one time point. Proteomic approaches have traditionally been used to measure only one state at a time because quantitation is difficult, especially when mass spectrometry is used as a readout. Isotopically labeled reagents have recently been introduced that allow comparison of two or three different states by mass spectrometry. Here, we evaluate the reproducibility of an experiment that measures three states simultaneously through stable isotope labeling of cells with amino acids in cell culture (SILAC) using light, medium, and heavy versions of amino acids. The major goal of this study was to assess the reproducibility of such experiments in combination with liquid chromatography tandem mass spectrometry (LC-MS/MS). Our results show that it is possible to obtain reproducible quantitative data to study protein dynamics based on our analysis of more than 220 peptide sets derived from. 20 proteins from 3 different LC-MS/MS runs.