Okada Ario, Gotoh Yasuhiro, Watanabe Takafumi, Furuta Eiji, Yamamoto Kaneyoshi, Utsumi Ryutaro
Department of Bioscience, Kinki University, Nara, Japan.
Methods Enzymol. 2007;422:386-95. doi: 10.1016/S0076-6879(06)22019-6.
We have developed two screening systems for isolating inhibitors that target bacterial two-component signal transduction: (1) a differential growth assay using a temperature-sensitive yycF mutant (CNM2000) of Bacillus subtilis, which is supersensitive to histidine kinase inhibitors, and (2) a high-throughput genetic system for targeting the homodimerization of histidine kinases essential for the bacterial two-component signal transduction. By using these methods, we have been able to identify various types of inhibitors that block the autophosphorylation of histidine kinases with different modes of actions.
我们开发了两种筛选系统，用于分离靶向细菌双组分信号转导的抑制剂：（1）一种差异生长测定法，使用枯草芽孢杆菌的温度敏感型yycF突变体（CNM2000），该突变体对组氨酸激酶抑制剂超敏感；（2）一种高通量遗传系统，用于靶向细菌双组分信号转导所必需的组氨酸激酶的同二聚化。通过使用这些方法，我们已经能够鉴定出各种类型的抑制剂，它们以不同的作用模式阻断组氨酸激酶的自磷酸化。
