An immuno-dot blot assay for screening histidine kinase inhibitors
An immuno-dot blot assay for screening histidine kinase inhibitors
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用于筛选组氨酸激酶抑制剂的免疫斑点印迹测定
DOI：
10.1016/j.ab.2020.113765
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发表时间：
2020
影响因子：
2.9
通讯作者：
Koike Tohru
中科院分区：
文献类型：
--
作者：
Kinoshita-Kikuta Emiko;Maruta Shino;Eguchi Yoko;Igarashi Masayuki;Okajima Toshihide;Utsumi Ryutaro;Kinoshita Eiji;Koike Tohru
由组氨酸激酶（HK）及其同源反应调节因子组成的双组分信号转导系统（TCSs）在细菌中普遍存在，并与病原体的毒力有关。tcs是替代抗生素和抗毒剂的潜在靶点。因此，测定细菌tcs中HK的活性是非常重要的。在这里，我们描述了一种使用抗n3 -磷酸组氨酸抗体的HKs抑制谱的免疫点印迹试验。这种简单的方法有望可靠地检测HK活性，并且可能适用于HK抑制剂的高通量筛选。
Two-component signal transduction systems (TCSs), consisting of a histidine kinase (HK) and its cognate response regulator, are ubiquitous among bacteria and are associated with the virulence of pathogens. TCSs are potential targets for alternative antibiotics and antivirulence agents. It is, thus, very important to determine HK activity in bacterial TCSs. Here, we describe an immuno-dot blot assay for the inhibition profiling of HKs using the anti-N3-phosphohistidine antibody. This simple method promises reliable detection of HK activity, and it is likely applicable in high-throughput screening of HK inhibitors.