The Center for Applied Proteomics and Molecular Medicine, George Mason University, Manassas, Virginia 20110, United States.
J Proteome Res. 2012 Apr 6;11(4):2127-39. doi: 10.1021/pr2007957. Epub 2012 Feb 27.
Liquid chromatography-tandem mass spectrometry (LC-MS/MS) and multiple reaction monitoring mass spectrometry (MRM-MS) proteomics analyses were performed on eccrine sweat of healthy controls, and the results were compared with those from individuals diagnosed with schizophrenia (SZ). This is the first large scale study of the sweat proteome. First, we performed LC-MS/MS on pooled SZ samples and pooled control samples for global proteomics analysis. Results revealed a high abundance of diverse proteins and peptides in eccrine sweat. Most of the proteins identified from sweat samples were found to be different than the most abundant proteins from serum, which indicates that eccrine sweat is not simply a plasma transudate and may thereby be a source of unique disease-associated biomolecules. A second independent set of patient and control sweat samples were analyzed by LC-MS/MS and spectral counting to determine qualitative protein differential abundances between the control and disease groups. Differential abundances of selected proteins, initially determined by spectral counting, were verified by MRM-MS analyses. Seventeen proteins showed a differential abundance of approximately 2-fold or greater between the SZ pooled sample and the control pooled sample. This study demonstrates the utility of LC-MS/MS and MRM-MS as a viable strategy for the discovery and verification of potential sweat protein disease biomarkers.
采用液相色谱-串联质谱(LC-MS/MS)和多重反应监测质谱(MRM-MS)蛋白质组学分析方法对健康对照者的外分泌汗液进行分析,并将结果与诊断为精神分裂症(SZ)个体的结果进行比较。这是首次对汗蛋白组进行大规模研究。首先,我们对 SZ 样本和对照样本进行 LC-MS/MS 分析,以进行全局蛋白质组学分析。结果显示,外分泌汗液中含有丰富多样的蛋白质和肽。从汗液样本中鉴定出的大多数蛋白质与血清中最丰富的蛋白质不同,这表明外分泌汗液不仅仅是一种血浆渗出物,因此可能是独特的与疾病相关生物分子的来源。第二组独立的患者和对照汗液样本通过 LC-MS/MS 和谱计数分析,以确定对照组和疾病组之间的定性蛋白质差异丰度。通过谱计数初步确定的选定蛋白质的差异丰度通过 MRM-MS 分析进行验证。在 SZ 混合样本和对照混合样本之间,有 17 种蛋白质的丰度差异约为 2 倍或更高。这项研究证明了 LC-MS/MS 和 MRM-MS 作为一种可行的策略,可用于发现和验证潜在的汗液蛋白疾病生物标志物。