A Two-Way Proteome Microarray Strategy to Identify Novel Mycobacterium tuberculosis-Human Interactors
A Two-Way Proteome Microarray Strategy to Identify Novel Mycobacterium tuberculosis-Human Interactors
复制标题
识别新型结核分枝杆菌-人类相互作用物的双向蛋白质组微阵列策略
DOI：
10.3389/fcimb.2019.00065
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发表时间：
2019-03-28
影响因子：
5.7
通讯作者：
Zhang, Zongde
中科院分区：
文献类型：
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作者：
Cao, Tingming;Lyu, Lingna;Zhang, Zongde
结核病(TB)是由结核分枝杆菌(Mtb)引起的严重威胁人类健康的疾病。未能消除结核病的主要原因是对结核分枝杆菌致病的分子机制缺乏清楚的了解。检测人类结核分枝杆菌相互作用蛋白使我们能够表征其机制，并确定结核病诊断和治疗的潜在分子靶点。然而，实验系统的Mtb相互作用装置并不是现成的。在这项研究中，我们实施了一种基于双向蛋白质组微阵列的无偏见的方法，系统地筛选全球人类Mtb相互作用因子，并确定Mtb效应因子的结合伙伴。我们的结果首次筛选出84个潜在的人类结核分枝杆菌相互作用因子。生物信息学分析进一步表明，这些候选蛋白可能参与了广泛的细胞功能，如激活DNA内源性启动子，转录DNA/RNA和坏死，以及免疫相关的信号通路。然后，利用Mtb蛋白质组芯片，结合His标签下拉实验和Co-IP，我们分别鉴定了候选蛋白NRF1的一个相互作用伙伴(Rv0577)和Smad2的三个结合伙伴(Rv0577、Rv2117、Rv2423)。这项研究为可能参与结核分枝杆菌发病机制的全球结核分枝杆菌相互作用因子的概况提供了新的见解，并展示了发现结核分枝杆菌效应因子的强大策略。
Tuberculosis (TB) is still a serious threat to human health which is caused by mycobacterium tuberculosis (Mtb). The main reason for failure to eliminate TB is lack of clearly understanding the molecular mechanism of Mtb pathogenesis. Determining human Mtb-interacting proteins enables us to characterize the mechanism and identify potential molecular targets for TB diagnosis and treatment. However, experimentally systematic Mtb interactors are not readily available. In this study, we performed an unbiased, comprehensive two-way proteome microarray based approach to systematically screen global human Mtb interactors and determine the binding partners of Mtb effectors. Our results, for the first time, screened 84 potential human Mtb interactors. Bioinformatic analysis further highlighted these protein candidates might engage in a wide range of cellular functions such as activation of DNA endogenous promoters, transcription of DNA/RNA and necrosis, as well as immune-related signaling pathways. Then, using Mtb proteome microarray followed His tagged pull-down assay and Co-IP, we identified one interacting partner (Rv0577) for the protein candidate NRF1 and three binding partners (Rv0577, Rv2117, Rv2423) for SMAD2, respectively. This study gives new insights into the profile of global Mtb interactors potentially involved in Mtb pathogenesis and demonstrates a powerful strategy in the discovery of Mtb effectors.