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首页> 外文期刊>International Journal of Biological Macromolecules: Structure, Function and Interactions >Predictive and fluorescent nanosensing experimental methods for evaluating anthrax protective antigen and lethal factor interactions for therapeutic applications
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Predictive and fluorescent nanosensing experimental methods for evaluating anthrax protective antigen and lethal factor interactions for therapeutic applications

机译:用于评估炭疽保护抗原和治疗应用致命因子相互作用的预测和荧光纳米溶解试验方法

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Recently, specific interaction of anthrax protective antigen domain 4 (PAD4) and lethal factor domain 1 (LFD1) have been considered for the design of novel diagnostic and therapeutic systems in medicine. In this study, theoretical and experimental approaches were used to monitor the interactions of PAD4 and LFD1. CLusPro server and Dimplot software were used to predict the interaction of these domains. Results, revealed interactive sites between PAD4 and LFD1 on loop regions of both C and N terminal of PAD4. In experimental methods, PAD4 and LFD1 were expressed in Escherichia coli and purified for usage in Magnetic Bead (MB) and Multi-Walled Carbon Nanotubes (MWCNTs) based bio-sensing platforms. In the magnetic-based system, the magnetic sedimentation of QD-PAD4 by MBs-LFD1 and the observation of the fluorescence spectrum related to QD-PAD4 in the precipitated materials confirmed the interaction of PAD4 with LFD1 protein. In the MWCNTs-based method, the QD-PAD4 fluorescence was quenched by absorption on MWCNTs. Upon the addition of LFD1, fluorescence emission was recovered, indicating interaction of LFD1 with QD-PAD4, which results the separation of QD-PAD4 from MWCNTs surfaces and fluorescence restoration. Finally, new approaches showed the interaction of PAD4 and LFD1, which can be used as an attractive model in medicine. (C) 2020 Elsevier B.V. All rights reserved.
机译:最近,已经考虑了炭疽保护抗原结构域4(PAD4)和致死因子结构域1(LFD1)的特异性相互作用用于设计新型诊断和治疗系统。在本研究中,使用理论和实验方法来监测PAD4和LFD1的相互作用。 cluspro服务器和Dimplot软件用于预测这些域的交互。结果,在PAD4的C和N端子的循环区域上揭示了PAD4和LFD1之间的交互点。在实验方法中,PAD4和LFD1在大肠杆菌中表达,并纯化用于磁珠(MB)和基于多壁碳纳米管(MWCNT)的生物传感平台的用途。在磁性系统中,QD-PAD4通过MBS-LFD1的磁沉降和沉淀材料中QD-PAD4相关的荧光光谱的观察证实了PAD4与LFD1蛋白的相互作用。在基于MWCNTS的方法中,通过在MWCNT上吸收QD-PAD4荧光淬灭。在添加LFD1时,回收荧光发射,表明LFD1与QD-PAD4的相互作用,这导致QD-PAD4与MWCNT表面和荧光复原的分离。最后,新方法显示了PAD4和LFD1的相互作用,可用作医学中的有吸引力的模型。 (c)2020 Elsevier B.v.保留所有权利。

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