Identification of astrocyte secreted proteins with a combination of shotgun proteomics and bioinformatics.
Identification of astrocyte secreted proteins with a combination of shotgun proteomics and bioinformatics.
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DOI：
10.1021/pr900248y
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发表时间：
2009-08
影响因子：
4.4
通讯作者：
Li L
中科院分区：
文献类型：
--
作者：
Dowell JA;Johnson JA;Li L
星形胶质细胞是哺乳动物正常脑功能的重要调节因子，包括突触信号传导、突触形成、神经元健康和存活。许多这些功能是通过分泌的蛋白质来执行的。为了分析星形胶质细胞分泌组，采用鸟枪蛋白质组学和生物信息学相结合的方法对小鼠星形胶质细胞原代培养的条件培养基进行了分析。利用二维和一维LC-MS/MS对星形胶质细胞分泌蛋白进行分析，鉴定出420多种蛋白。为了完善我们的结果，使用细胞质控制去除细胞内蛋白质污染物。在进一步的细化中，假定的分泌蛋白被SignalP、SecretomeP和基因本体分析所分析，得到187种分泌蛋白的细化列表。总之，使用散弹枪蛋白质组学结合多轮数据细化产生了高质量的星形胶质细胞分泌蛋白目录。
Astrocytes are important regulators of normal brain function in mammals, including roles in synaptic signaling, synapse formation, and neuronal health and survival. Many of these functions are executed via secreted proteins. In order to analyze the astrocyte secretome, a combination of shotgun proteomics and bioinformatics was employed to analyze conditioned media from primary murine astrocyte cultures. Both two- and one-dimensional LC-MS/MS were used to analyze astrocyte secreted proteins, resulting in the identification of over 420 proteins. In order to refine our results, the intracellular protein contaminants were removed in silico using a cytoplasmic control. In additional rounds of refinement, putative secreted proteins were subjected to analysis by SignalP, SecretomeP, and gene ontology analysis, yielding a refined list of 187 secreted proteins. In conclusion, the use of shotgun proteomics combined with multiple rounds of data refinement produced a high quality catalog of astrocyte secreted proteins.
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