过表达sigma-1受体对ADAM17和ADAM10功能的抑制作用研究

发布时间：2018-12-12 10:53

【摘要】：Sigma受体是一种内质网分子伴侣蛋白,它可以结合多类配体,并广泛参与了脂类代谢及转运、神经突发生、离子通道、细胞存活等调控过程,从而与人类神经系统疾病(如ALS、阿尔茨海默病、抑郁)以及中风、癌症等许多疾病都息息相关。ALS是一种运动神经元退化引起的致命神经退行性疾病。在ALS-FTD/FTD家族中,对SIGMAR1基因进行测序发现一个错位突变,突变影响定位在跨膜域的高度保守氨基酸,而该基因编码蛋白即sigma-1受体。然而,目前还不清楚这个突变如何影响运动神经元细胞功能。 ADAM 17和ADAM 10是一类膜锚定的金属蛋白酶家族的成员,加工和剪切生长因子、细胞因子配体和受体、细胞粘着因子及Notch-1等多种膜锚定蛋白的胞外结构域,因而与诸如炎症、癌症、阿尔茨海默病等多种疾病的病理过程相关。目前,有报道ADAM 10和ADAM 17的功能均与其在脂筏区和非脂筏区的分布相关,然而sigma-1受体是否影响ADAM17/10功能的行使还不清楚。Sigma-1受体参与了内质网脂质转运,上调sigma-1受体会改变脂筏中脂质部分而影响脂筏。因其介导质膜中脂筏重组,从而可能影响存在于质膜脂筏中蛋白的功能。本实验通过碱性磷酸酶标签底物的报告系统发现,在不同细胞系上过表达sigma-1受体,分别抑制了ADAM17、ADAM10依赖的底物HB-EGF及BTC的剪切释放作用,而两者蛋白表达水平无明显变化。细胞免疫荧光实验发现,过表达sigma-1受体后脂筏标志蛋白在细胞上的分布发生变化。
[Abstract]:Sigma receptor is an endoplasmic reticulum molecular chaperone protein that binds to multiple ligands and is widely involved in lipid metabolism and transport, neuronogenesis, ion channels, cell survival and other regulatory processes, thus associated with human nervous system diseases (such as ALS,) Alzheimer's disease, depression) and stroke, cancer and many other diseases are linked. ALS is a fatal neurodegenerative disease caused by motor neuron degeneration. In the ALS-FTD/FTD family, the SIGMAR1 gene was sequenced and a translocation mutation was found, which affected the highly conserved amino acids located in the transmembrane domain, and the protein encoded by the gene was the sigma-1 receptor. However, it is not clear how this mutation affects the function of motor neuron cells. ADAM 17 and ADAM 10 are members of a family of membrane-anchored metalloproteinases, processing and shearing growth factors, cytokine ligands and receptors, cell adhesion factors and Notch-1, the extracellular domains of many membrane anchored proteins, and thus associated with inflammation, Cancer, Alzheimer's disease and other diseases are related to the pathological process. At present, it has been reported that the functions of ADAM 10 and ADAM 17 are related to their distribution in lipid raft and non-fat raft. However, it is not clear whether sigma-1 receptor affects the function of ADAM17/10. Sigma-1 receptor is involved in endoplasmic reticulum lipid transport. Upregulation of sigma-1 receptor affects lipid rafts by altering the lipid portion of the lipid rafts. It may affect the function of protein in plasma membrane lipid raft because it mediates the recombination of lipid raft in plasma membrane. In this study, we found that the expression of sigma-1 receptors in different cell lines inhibited the shear release of ADAM17,ADAM10 dependent substrates HB-EGF and BTC, respectively. The expression level of both proteins did not change significantly. Cell immunofluorescence assay showed that the distribution of lipid raft marker protein changed after overexpression of sigma-1 receptor.
【学位授予单位】：复旦大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R363

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