Proteomics Analysis Reveals Overlapping Functions of Clustered Protocadherins
Proteomics Analysis Reveals Overlapping Functions of Clustered Protocadherins
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DOI：
10.1074/mcp.m900343-mcp200
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发表时间：
2010-01-01
影响因子：
7
通讯作者：
Wang, Xiaozhong
中科院分区：
文献类型：
--
作者：
Han, Meng-Hsuan;Lin, Chengyi;Wang, Xiaozhong
原钙粘蛋白（Protocadherin，Pcdh）基因簇Pcdh-α、Pcdh-β和Pcdh-γ在脊椎动物中枢神经系统发育中起重要作用。为了深入了解PCDH的分子作用，我们对PCDH-γ相关蛋白复合物进行了系统的蛋白质组学分析。我们确定了154个非冗余蛋白质的PCDH-γ复合物的列表。该列表包括近30个成员的集群Pcdh-α，β和γ家族的核心组成部分的复合物和另外超过120个推定的PCDH相关蛋白。我们使用特异性抗体验证了PCDH-γ相关蛋白的选定子集。PCDH相关蛋白的身份分析表明，他们中的大多数重叠与突触后密度制剂的蛋白质组学概况。膜蛋白复合物的进一步分析显示，几个验证的PCDH-γ相关蛋白表现出减少的水平在Pcdh-γ缺陷的脑组织。因此，PCDH-γ是复合物完整性所必需的。然而，整体复合物的大小和许多其他蛋白质的丰度保持不变，提高了PCDH-α和PCDH-β在复合物形成中可能补偿PCDH-γ功能的可能性。作为这一想法的测试，PCDH-α s和PCDH-γ s的RNA干扰敲低表明，PCDH在调节鸡脊髓神经元存活方面具有冗余功能。总之，我们的数据提供的证据表明，集群PCDH共存于大型蛋白质复合物，并在脊椎动物神经发育过程中有重叠的功能。Molecular & Cellular Proteomics 9：71-83，2010.
The three tandem-arrayed protocadherin (Pcdh) gene clusters, namely Pcdh-alpha, Pcdh-beta, and Pcdh-gamma, play important roles in the development of the vertebrate central nervous system. To gain insight into the molecular action of PCDHs, we performed a systematic proteomics analysis of PCDH-gamma-associated protein complexes. We identified a list of 154 non-redundant proteins in the PCDH-gamma complexes. This list includes nearly 30 members of clustered Pcdh-alpha, -beta, and -gamma families as core components of the complexes and additionally over 120 putative PCDH-associated proteins. We validated a selected subset of PCDH-gamma-associated proteins using specific antibodies. Analysis of the identities of PCDH-associated proteins showed that the majority of them overlap with the proteomic profile of postsynaptic density preparations. Further analysis of membrane protein complexes revealed that several validated PCDH-gamma-associated proteins exhibit reduced levels in Pcdh-gamma-deficient brain tissues. Therefore, PCDH-gamma s are required for the integrity of the complexes. However, the size of the overall complexes and the abundance of many other proteins remained unchanged, raising a possibility that PCDH-alpha s and PCDH-beta s might compensate for PCDH-gamma function in complex formation. As a test of this idea, RNA interference knockdown of both PCDH-alpha s and PCDH-gamma s showed that PCDHs have redundant functions in regulating neuronal survival in the chicken spinal cord. Taken together, our data provide evidence that clustered PCDHs coexist in large protein complexes and have overlapping functions during vertebrate neural development. Molecular & Cellular Proteomics 9: 71-83, 2010.