McKusick-Nathans Institute of Genetic Medicine, Johns Hopkins University, Baltimore, Maryland 21205, USA.
J Proteomics. 2011 Mar 1;74(3):294-303. doi: 10.1016/j.jprot.2010.11.006. Epub 2010 Nov 13.
Signal transduction pathways are tightly controlled by positive and negative regulators. We have previously identified Odin (also known as ankyrin repeat and sterile alpha motif domain-containing 1A; gene symbol ANKS1A) as a negative regulator of growth factor signaling; however, the mechanisms through which Odin regulates these pathways remain to be elucidated. To determine how Odin negatively regulates growth factor signaling, we undertook a proteomic approach to systematically identify proteins that interact with Odin using the SILAC strategy. In this study, we identified 18 molecules that were specifically associated in a protein complex with Odin. Our study established that the complete family of 14-3-3 proteins occur in a protein complex with Odin, which is also supported by earlier reports that identified a few members of the 14-3-3 family as Odin interactors. Among the novel protein interactors of Odin were CD2-associated protein, SH3 domain kinase binding protein 1 and DAB2 interacting protein. We confirmed 8 of the eighteen interactions identified in the Odin protein complex by co-immunoprecipitation experiments. Finally, a literature-based network analysis revealed that Odin interacting partners are involved in various cellular processes, some of which are key molecules in regulating receptor endocytosis.
信号转导途径受到正、负调节剂的严密控制。我们先前已确定 Odin（也称为锚蛋白重复和无菌α基序域 1A；基因符号为 ANKS1A）是生长因子信号的负调节剂；但是，Odin 调节这些途径的机制仍有待阐明。为了确定 Odin 如何负调控生长因子信号，我们采用了一种蛋白质组学方法，使用 SILAC 策略系统地鉴定与 Odin 相互作用的蛋白质。在这项研究中，我们鉴定了 18 种分子，这些分子与 Odin 特异性地存在于一个蛋白质复合物中。我们的研究确定了完整的 14-3-3 蛋白家族与 Odin 发生相互作用，这也得到了先前报道的支持，该报道鉴定了 14-3-3 家族的一些成员为 Odin 相互作用子。在 Odin 的新型蛋白质相互作用子中包括 CD2 相关蛋白、SH3 结构域激酶结合蛋白 1 和 DAB2 相互作用蛋白。我们通过共免疫沉淀实验验证了 Odin 蛋白复合物中鉴定出的 18 个相互作用中的 8 个。最后，基于文献的网络分析表明，Odin 相互作用伙伴参与了各种细胞过程，其中一些是调节受体内吞作用的关键分子。
