Key Laboratory for Rare Disease Research of Shandong Province, Key Laboratory for Biotech Drugs of the Ministry of Health, Shandong Medical Biotechnological Center, Shandong Academy of Medical Sciences, Ji'nan, Shandong, China.
Biosci Trends. 2013 Jun;7(3):144-51.
Matrix vesicles (MVs) involved in the initiation of mineralization by deposition of hydroxyapatite (HA) in their lumen are released by the budding of mineralization-competent cells during skeletogenesis and bone development. To identify additional mineralization-related proteins, MVs were isolated from non-stimulated and stimulated Saos-2 cells in culture via an Exoquick™ approach and the corresponding proteomes were identified and quantified with label-free quantitative proteome technology. The isolated MVs were confirmed by electron microscopy, alkaline phosphatase activity (ALP), biomarkers, and mineral formation analyses. Label-free quantitative proteome analysis revealed that 19 calcium binding proteins (CaBPs), including Grp94, calnexin, calreticulin, calmodulin, and S100A4/A10, were up-regulated in MVs of Saos-2 cells upon stimulation of mineralization. This result provides new clues to study the mechanism of the initiation of MV-mediated mineralization.
基质小泡 (MVs) 通过在腔室内沉积羟基磷灰石 (HA) 参与矿化的启动，在成骨过程中通过矿化能力细胞的出芽释放。为了鉴定其他与矿化相关的蛋白质，通过 Exoquick™ 方法从非刺激和刺激的 Saos-2 细胞中分离 MV，并使用无标记定量蛋白质组学技术鉴定和定量相应的蛋白质组。通过电子显微镜、碱性磷酸酶活性 (ALP)、生物标志物和矿化形成分析证实了分离的 MV。无标记定量蛋白质组学分析显示，在矿化刺激下，Saos-2 细胞的 MV 中上调了 19 种钙结合蛋白 (CaBP)，包括 Grp94、钙连蛋白、钙网蛋白、钙调蛋白和 S100A4/A10。这一结果为研究 MV 介导的矿化起始的机制提供了新的线索。
