Dumont Debora, Noben Jean-Paul, Verhaert Peter, Stinissen Piet, Robben Johan
Hasselt University, Biomedical Research Institute and transnationale Universiteit Limburg, School of Life Sciences, Diepenbeek, Belgium.
Proteomics. 2006 Sep;6(18):4967-77. doi: 10.1002/pmic.200600080.
This paper reports on the findings of the Biomedical Research Institute, as one of the participants in the pilot study of the HUPO Brain Proteome Project. A biopsy and autopsy study sample derived from human brain was distributed among the participants for proteomic analysis. In our laboratory, attention was focused on protein identification using the bottom-up shotgun approach. Protein extracts derived from both samples were trypsinized and analyzed separately by 2-D LC and MS. In a complementary approach, the tryptic digests were analyzed directly by LC-ESI-MS/MS and gas-phase fractionation in the mass spectrometer. Taken together, both proteomic approaches in combination with a stringent evaluation process, resulted in the confident identification of 209 proteins in the human brain samples under investigation.
本文报告了生物医学研究所作为人类蛋白质组组织（HUPO）脑蛋白质组计划试点研究参与者之一的研究结果。取自人脑的活检和尸检研究样本分发给各参与者进行蛋白质组分析。在我们实验室，重点是使用自下而上的鸟枪法进行蛋白质鉴定。从两个样本中提取的蛋白质经胰蛋白酶消化后，分别通过二维液相色谱和质谱进行分析。作为一种补充方法，胰蛋白酶消化产物通过液相色谱-电喷雾串联质谱（LC-ESI-MS/MS）和质谱仪中的气相分级分离直接进行分析。综合起来，这两种蛋白质组学方法与严格的评估过程相结合，使得在所研究的人脑样本中可靠地鉴定出了209种蛋白质。
