Differential protein expression in male and female human lumbar cerebrospinal fluid using iTRAQ reagents after abundant protein depletion
Differential protein expression in male and female human lumbar cerebrospinal fluid using iTRAQ reagents after abundant protein depletion
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DOI：
10.1002/pmic.200700455
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发表时间：
2007-10-01
期刊：
影响因子：
3.4
通讯作者：
Muddiman, David C.
中科院分区：
文献类型：
--
作者：
Ogata, Yuko;Charlesworth, M. Cristine;Muddiman, David C.
脑脊液（CSF）已成为最常用的生物介质之一。对于神经系统疾病的生理学研究，由于其接近大脑和临床可用性;然而，在采取合理的方法进行生物标志物发现或诊断之前，了解亚群中CSF蛋白质组的基本特征至关重要。在这项研究中，我们研究了蛋白质的差异表达，在合并的男性和女性的CSF利用同量异位素标签的相对和绝对定量。（iTRAQ（TM））试剂在使用多重亲和去除系统（MARS）去除六种高丰度蛋白质后的测定。共鉴定出219种蛋白质（95%置信水平），其中12种蛋白质的表达水平存在差异。12个差异表达蛋白中有11个显示雄性/雌性比值在1.15和1.29之间（重复平均值），表明雄性和雌性CSF之间具有显著的相似性。一个显著的例外是雄性CSF中血浆铜蓝蛋白（铁氧化酶）的表达水平略低（0.81），这是一种含铜蛋白，可催化亚铁转化为三价铁，具有抗氧化特性。我们还使用Western印迹分析检查了构成合并CSF的每个个体患者样品中的铜蓝蛋白水平，其证实了雄性CSF中铜蓝蛋白的较低表达水平。
Cerebrospinal fluid (CSF) has become one of the most frequently used biological medium. for physiological studies for neurological disorders due to its proximity to the brain and clinical availability; however, before undertaking a rational approach to biomarker discovery or diagnostics, it is crucial to understand the underlying characteristics of CSF proteome in subpopulations. In this study, we examined the differential expression of proteins in pooled male and female CSF utilizing isobaric tags for relative and absolute quantification. (iTRAQ(TM)) reagents after the depletion of six high abundant proteins using a multiple affinity removal system (MARS). A total of 219 proteins were identified (95% confidence level), and 12 proteins showed difference in expression levels. Eleven out of 12 differentially expressed proteins showed ratios of male/female between 1.15 and 1.29 (duplicate average), indicating a remarkable similarity between male and female CSF. One notable exception was the slightly lower expression level of ceruloplasmin (ferroxidase) in male CSF (0.81), a copper containing protein that catalyzes the conversion of ferrous iron to ferric iron with antioxidant properties. We also examined the levels of ceruloplasmin in each individual patient sample which constituted the pooled CSF using Western blot analysis which confirmed the lower expression levels of ceruloplasmin in male CSF.