Burgess Jennifer A, Lescuyer Pierre, Hainard Alexandre, Burkhard Pierre R, Turck Natacha, Michel Philippe, Rossier Joël S, Reymond Frédéric, Hochstrasser Denis F, Sanchez Jean-Charles
Biomedical Proteomics Research Group, Department of Structural Biology and Bioinformatics, Faculty of Medicine, University of Geneva, Geneva, Switzerland.
J Proteome Res. 2006 Jul;5(7):1674-81. doi: 10.1021/pr060160v.
Following any form of brain insult, proteins are released from damaged tissues into the cerebrospinal fluid (CSF). This body fluid is therefore an ideal sample to use in the search for biomarkers of neurodegenerative disorders and brain damage. In this study, we used human post-mortem CSF as a model of massive brain injury and cell death for the identification of such protein markers. Pooled post-mortem CSF samples were analyzed using a protocol that combined immunoaffinity depletion of abundant CSF proteins, off-gel electrophoresis, SDS-PAGE and protein identification by LC-MS/MS. A total of 299 proteins were identified, of which 172 proteins were not previously described to be present in CSF. Of these 172 proteins, more than 75% have been described as intracellular proteins suggesting that they were released from damaged cells. Immunoblots of a number of proteins were performed on individual post-mortem CSF samples and confirmed elevated concentrations in post-mortem CSF compared to ante-mortem CSF. Interestingly, among the proteins specifically identified in the post-mortem CSF, several have been previously described as biochemical markers of brain damage.
在遭受任何形式的脑损伤后，蛋白质会从受损组织释放到脑脊液（CSF）中。因此，这种体液是寻找神经退行性疾病和脑损伤生物标志物的理想样本。在本研究中，我们使用人类尸检脑脊液作为大规模脑损伤和细胞死亡的模型，以鉴定此类蛋白质标志物。使用一种结合了对脑脊液中丰富蛋白质进行免疫亲和去除、离胶电泳、SDS-PAGE以及通过LC-MS/MS进行蛋白质鉴定的方案，对汇集的尸检脑脊液样本进行分析。总共鉴定出299种蛋白质，其中172种蛋白质此前未被描述存在于脑脊液中。在这172种蛋白质中，超过75%被描述为细胞内蛋白质，这表明它们是从受损细胞中释放出来的。对多个个体尸检脑脊液样本进行了多种蛋白质的免疫印迹分析，结果证实与生前脑脊液相比，尸检脑脊液中的浓度升高。有趣的是，在尸检脑脊液中特异性鉴定出的蛋白质中，有几种此前已被描述为脑损伤的生化标志物。
