Enhanced analysis of metastatic prostate cancer using stable isotopes and high mass accuracy instrumentation
Enhanced analysis of metastatic prostate cancer using stable isotopes and high mass accuracy instrumentation
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DOI：
10.1021/pr0504891
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发表时间：
2006-05-01
影响因子：
4.4
通讯作者：
Gygi, SP
中科院分区：
文献类型：
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作者：
Everley, PA;Bakalarski, CE;Gygi, SP
蛋白质组学的主要目标是更好地了解蛋白质表达水平的生物学功能。随着该领域的成熟，许多技术正在开发中，以帮助在近全球范围内识别、量化和表征蛋白质表达和翻译后修饰。细胞培养中氨基酸的稳定同位素标记就是这样一种技术，已显示出广泛的生物学应用。虽然我们最近展示了该技术在转移性前列腺癌模型中的应用，但我们现在报告使用线性离子阱傅里叶变换离子回旋共振质谱仪 (LTQ FT) 和新颖的定量软件在定量分析方面取得了重大改进。这导致了近 1400 种蛋白质的定量，与我们之前的研究相比增加了 3 倍以上。蛋白质组覆盖率的显着增加可归因于 (1) 使用双标记策略，(2) LTQ FT 质谱仪提供更高的灵敏度、速度和质量准确度，以及 (3) 更强大的定量软件。最后，通过使用串联的目标/诱饵蛋白数据库进行肽搜索，我们现在在估计假阳性率为百分之一的情况下报告这些数据。
The primary goal of proteomics is to gain a better understanding of biological function at the protein expression level. As the field matures, numerous technologies are being developed to aid in the identification, quantification and characterization of protein expression and post-translational modifications on a near-global scale. Stable isotope labeling by amino acids in cell culture is one such technique that has shown broad biological applications. While we have recently shown the application of this technology to a model of metastatic prostate cancer, we now report a substantial improvement in quantitative analysis using a linear ion-trap Fourier transform ion cyclotron resonance mass spectrometer (LTQ FT) and novel quantification software. This resulted in the quantification of nearly 1400 proteins, a greater than 3-fold increase in comparison to our earlier study. This dramatic increase in proteome coverage can be attributed to (1) use of a double-labeling strategy, (2) greater sensitivity, speed and mass accuracy provided by the LTQ FT mass spectrometer, and (3) more robust quantification software. Finally, by using a concatenated target/decoy protein database for our peptide searches, we now report these data in the context of an estimated false-positive rate of one percent.