A Single Tool to Monitor Multiple Protein-Protein Interactions of the Escherichia coli Acyl Carrier Protein
A Single Tool to Monitor Multiple Protein-Protein Interactions of the Escherichia coli Acyl Carrier Protein
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DOI：
10.1021/acsinfecdis.9b00150
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发表时间：
2019-09-01
影响因子：
5.3
通讯作者：
Burkart, Michael D.
中科院分区：
文献类型：
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作者：
Charov, Katherine;Burkart, Michael D.
蛋白质之间的相互作用在生命的各个领域都是普遍存在的，最近作为药物靶点引起了人们的兴趣。然而，目前许多研究蛋白质-蛋白质相互作用的方法都是昂贵且低通量的。在这里，我们展示了一种基于大肠杆菌酰基载体蛋白（EcACP）的自然翻译后修饰的溶剂化变色工具，用于可视化EcACP与来自几种生物合成途径的13种不同伙伴酶之间的蛋白质-蛋白质相互作用。我们使用这个工具来确认EcACP与催化和调节蛋白之间的相互作用。我们还展示了这种方法在检测伙伴酶结构的变构变化和活性位点抑制剂的有效性方面的效用。我们预计未来将这种检测方法应用到抗生素发现的高通量筛选中。
Protein-protein interactions are ubiquitous to all domains of life and have gained recent interest as drug targets. However, many current methods to study protein-protein interactions can be costly and are low-throughput. Here, we demonstrate a solvatochromic tool based on the natural post-translational modification of the Escherichia coli acyl carrier protein (EcACP) used to visualize protein-protein interactions between EcACP and 13 different partner enzymes from several biosynthetic pathways. We use this tool to confirm proposed interactions between EcACP and both catalytic and regulatory proteins. We also show the utility of this method toward detecting allosteric changes to partner enzyme structure and the validation of active site inhibitors. We anticipate the future adaptation of this assay into a high-throughput screen for antibiotic discovery.