Univ Lille Nord de France, F-59000 Lille, France.
Proteome Sci. 2010 Nov 15;8:57. doi: 10.1186/1477-5956-8-57.
Brain capillary endothelial cells (BCECs) form the physiological basis of the blood-brain barrier (BBB). The barrier function is (at least in part) due to well-known proteins such as transporters, tight junctions and metabolic barrier proteins (e.g. monoamine oxidase, gamma glutamyltranspeptidase and P-glycoprotein). Our previous 2-dimensional gel proteome analysis had identified a large number of proteins and revealed the major role of dynamic cytoskeletal remodelling in the differentiation of bovine BCECs. The aim of the present study was to elaborate a reference proteome of Triton X-100-soluble species from bovine BCECs cultured in the well-established in vitro BBB model developed in our laboratory.
A total of 215 protein spots (corresponding to 130 distinct proteins) were identified by 2-dimensional gel electrophoresis, whereas over 350 proteins were identified by a shotgun approach. We classified around 430 distinct proteins expressed by bovine BCECs. Our large-scale gene expression analysis enabled the correction of mistakes referenced into protein databases (e.g. bovine vinculin) and constitutes valuable evidence for predictions based on genome annotation.
Elaboration of a reference proteome constitutes the first step in creating a gene expression database dedicated to capillary endothelial cells displaying BBB characteristics. It improves of our knowledge of the BBB and the key proteins in cell structures, cytoskeleton organization, metabolism, detoxification and drug resistance. Moreover, our results emphasize the need for both appropriate experimental design and correct interpretation of proteome datasets.
脑毛细血管内皮细胞（BCEC）构成了血脑屏障（BBB）的生理基础。该屏障功能（至少部分是由于）归因于众所周知的蛋白质，如转运蛋白、紧密连接和代谢屏障蛋白（如单胺氧化酶、γ-谷氨酰转肽酶和 P-糖蛋白）。我们之前的 2 维凝胶蛋白质组分析已经鉴定了大量的蛋白质，并揭示了动态细胞骨架重塑在牛 BCEC 分化中的主要作用。本研究的目的是详细阐述我们实验室建立的体外 BBB 模型中培养的牛 BCEC 中 Triton X-100 可溶物种的参考蛋白质组。
通过 2 维凝胶电泳鉴定了 215 个蛋白点（对应于 130 种不同的蛋白质），而通过鸟枪法鉴定了超过 350 种蛋白质。我们对牛 BCEC 表达的约 430 种不同蛋白质进行了分类。我们的大规模基因表达分析使我们能够纠正蛋白质数据库中的错误（例如牛纽蛋白），并为基于基因组注释的预测提供了有价值的证据。
参考蛋白质组的编制是创建专门针对具有 BBB 特征的毛细血管内皮细胞的基因表达数据库的第一步。它提高了我们对 BBB 的认识，以及对细胞结构、细胞骨架组织、代谢、解毒和耐药性的关键蛋白质的认识。此外，我们的结果强调了需要适当的实验设计和对蛋白质组数据集的正确解释。
