神经接头蛋白Dok6下游作用蛋白的鉴定及初步功能分析

Screening and potential functional analysis of substrates of neuronal adapter protein Dok6

目的 Dok6下游作用蛋白的鉴定和初步功能分析。方法利用anti-Dok6的抗体进行免疫共沉淀实验,然后通过Western blot,银染分析,质谱鉴定以及后续的生物信息学预测及分析其潜在的生理功能。结果本实验得到了26个可能与Dok6发生直接或间接相互作用的蛋白,这些蛋白分别参与了体内众多的生理过程。为了缩小研究范围,选取了与神经系统发育及细胞内信号传导相关的9个蛋白作为研究对象进行后续的功能研究。结论通过质谱鉴定以及生物信息学综合分析得到的潜在Dok6下游蛋白将给研究Dok6的生理功能及阐述其相关分子机制提供重要的依据。

Objective To screen the protein which can interact with Dok6 and explore the potential function. Methods Immunoprecipitation, western blot, and sliver staining were employed to screen the candidate proteins interact with Dok6. liquid chromatography-tandem mass spectrometry （LC-MS/MS） was used to identify the binding proteins of Dok6. Finally, we analyzed its potential physiological functions by bioinforrnatics prediction. Results After screening the binding proteins of Dok6 with IP and LC-MS/MS, 26 candidate binding proteins were identi fied. These proteins are involved in many physiological processes in the body. In order to narrow the scope of the study, we selected nine proteins associated with nervous system development and intracellular signal transduction as object of study for following study of the function. Conclusions The potential Dok6 downstream proteins identified by mass spectrometry and bioinformatics will provide some clues to determine physiological functions and related molecular mechanisms of Dok6.