Dagley Laura F, White Carl A, Liao Yang, Shi Wei, Smyth Gordon K, Orian Jacqueline M, Emili Andrew, Purcell Anthony W
Department of Biochemistry and Molecular Biology, Bio21 Molecular Science and Biotechnology Institute, University of Melbourne, Parkville, Victoria, Australia; Banting and Best Department of Medical Research, Terrence Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, ON, Canada; Department of Biochemistry and Molecular Biology, Monash University, Clayton, Victoria, Australia.
Proteomics. 2014 Feb;14(2-3):241-61. doi: 10.1002/pmic.201300196. Epub 2014 Jan 6.
Despite major advances in neuroscience, a comprehensive understanding of the structural and functional components of the adult brain compartments remains to be fully elucidated at a quantitative molecular level. Indeed, over half of the soluble- and membrane-annotated proteins are currently unmapped within online digital brain atlases. In this study, two complementary approaches were used to assess the unique repertoire of proteins enriched within select regions of the adult mouse CNS, including the brain stem, cerebellum, and remaining brain hemispheres. Of the 1200 proteins visualized by 2D-DIGE, approximately 150 (including cytosolic and membrane proteins) were found to exhibit statistically significant changes in relative abundance thus representing putative region-specific brain markers. In addition to using a high-precision (18) O-labeling strategy for the quantitative LC-MS/MS mapping of membrane proteins isolated from myelin-enriched fractions, we have identified over 1000 proteins that have yet to be described in any other mammalian myelin proteome. A comparison of our myelin proteome was made to an existing transcriptome database containing mRNA abundance profiles during oligodendrocyte differentiation and has confirmed statistically significant abundance changes for ∼500 of these newly mapped proteins, thus revealing new roles in oligodendrocyte and myelin biology. These data offer a resource for the neuroscience community studying the molecular basis for specialized neuronal activities in the CNS and myelin-related disorders. The MS proteomics data associated with this manuscript have been deposited to the ProteomeXchange Consortium with the dataset identifier PXD000327 (http://proteomecentral.proteomexchange.org/dataset/PXD000327).
尽管神经科学取得了重大进展，但在定量分子水平上，对成人大脑各部分的结构和功能成分的全面理解仍有待充分阐明。事实上，目前超过一半的可溶性和膜注释蛋白在在线数字脑图谱中尚未被定位。在本研究中，我们采用了两种互补的方法来评估成年小鼠中枢神经系统特定区域（包括脑干、小脑和其余脑半球）中富集的独特蛋白质库。通过二维差异凝胶电泳（2D-DIGE）可视化的1200种蛋白质中，约150种（包括胞质蛋白和膜蛋白）被发现其相对丰度有统计学上的显著变化，因此代表了假定的区域特异性脑标记物。除了使用高精度的（18）O标记策略对从富含髓磷脂的组分中分离的膜蛋白进行定量液相色谱-串联质谱（LC-MS/MS）图谱分析外，我们还鉴定出了1000多种在任何其他哺乳动物髓磷脂蛋白质组中尚未被描述的蛋白质。我们将髓磷脂蛋白质组与一个包含少突胶质细胞分化过程中mRNA丰度图谱的现有转录组数据库进行了比较，证实了这些新定位的蛋白质中约500种在统计学上有显著的丰度变化，从而揭示了它们在少突胶质细胞和髓磷脂生物学中的新作用。这些数据为神经科学界研究中枢神经系统中特殊神经元活动的分子基础以及与髓磷脂相关的疾病提供了资源。与本手稿相关的质谱蛋白质组学数据已存入蛋白质组交换联盟，数据集标识符为PXD000327（http://proteomecentral.proteomexchange.org/dataset/PXD000327）。
