Quantitative analysis of the low molecular weight serum proteome using 18O stable isotope labeling in a lung tumor xenograft mouse model
Quantitative analysis of the low molecular weight serum proteome using 18O stable isotope labeling in a lung tumor xenograft mouse model
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DOI：
10.1016/j.jasms.2005.02.005
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发表时间：
2005-08-01
影响因子：
3.2
通讯作者：
Karsan, A
中科院分区：
文献类型：
--
作者：
Hood, BL;Lucas, DA;Karsan, A
随着蛋白质组学分析技术和方法的进步，在生物流体如血清和血浆中发现生物标志物引起了极大的兴趣。目前的假设表明，低分子量（LMW）血清蛋白质组拥有一个档案的剪切和切割的蛋白质片段，可以提供深入了解疾病的发展。虽然这些生物流体代表了有吸引力的样本，可以从中发现新的和更准确的疾病生物标志物，但这些样本中固有的人与人之间的差异使它们的发现变得复杂。小鼠是研究人类疾病最广泛使用的动物模型之一，因为它们代表了高度可控的实验模型系统。在这项研究中，低分子量血清蛋白质组进行了比较异种移植肿瘤荷瘤小鼠和对照小鼠之间的差异标记利用胰蛋白酶介导的掺入稳定同位素的氧，O-18。将这些蛋白质状态合并，通过强阳离子交换色谱法进行分级分离，并通过纳米流反相液相色谱法与串联质谱法在线耦合进行分析，从而鉴定出至少由单个完全胰蛋白酶肽鉴定的6003种蛋白质。近1650蛋白质被确定和定量的两个或两个以上的完全胰蛋白酶肽。在这项工作中采用的方法提供了未来的定量测量比较动物模型的疾病和人类疾病队列的手段。
With advancements in the analytical technologies and methodologies in proteomics, there is great interest in biomarker discovery in biofluids such as serum and plasma. Current hypotheses suggest that the low molecular weight (LMW) serum proteome possesses an archive of clipped and cleaved protein fragments that may provide insight into disease development. Though these biofluids represent attractive samples from which new and more accurate disease biomarkers may be found, the intrinsic person-to-person variability in these samples complicates their discovery. Mice are one of the most extensively used animal models for studying human disease because they represent a highly controllable experimental model system. In this study, the LMW serum proteome was compared between xenografted tumor-bearing mice and control mice by differential labeling utilizing trypsin-mediated incorporation of the stable isotope of oxygen, O-18. The digestates were combined, fractionated by strong cation exchange chromatography, and analyzed by nanoflow reversed-phase liquid chromatography coupled online with tandem mass spectrometry, resulting in the identification of 6003 proteins identified by at least a single, fully tryptic peptide. Almost 1650 proteins were identified and quantitated by two or more fully tryptic peptides. The methodology adopted in this work provides the means for future quantitative measurements in comparative animal models of disease and in human disease cohorts.