The PilF protein from the thermophilic bacterium Thermus thermophilus is a traffic ATPase powering the assembly of the DNA translocation machinery as well as of type 4 pili. Thereby PilF mediates the natural transformability of T. thermophilus. PilF contains a C-terminal ATPase domain and three N-terminal domains with partial homology to so-called general secretory pathway II (GSPII) domains. These three GSPII domains (GSPII-A, GSPII-B and GSPII-C) are essential for pilus assembly and twitching motility. They show varying degrees of sequence homology to the N-terminal domain of the ATPase MshE from Vibrio cholerae which binds the bacterial second messenger molecule c-di-GMP. NMR experiments demonstrate that the GSPII-B domain of PilF also binds c-di-GMP with high affinity and forms a 1:1 complex in slow exchange on the NMR time scale. As a prerequisite for structural studies of c-di-GMP binding to the GSPII-B domain of T. thermophilus PilF we present here the NMR resonance assignments for the apo and the c-di-GMP bound state of GSPII-B. In addition, we map the binding site for c-di-GMP on the GSPII-B domain using chemical shift perturbation data and compare the dynamics of the apo and the c-di-GMP-bound state of the GSPII-B domain based on {¹H},¹⁵N-hetNOE data.

中文翻译：

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在载脂蛋白和c-di-GMP结合状态下，嗜热栖热菌的运输ATPase PilF的GSPII-B结构域的NMR共振分配。

来自嗜热嗜热菌Thermus thermophilus的PilF蛋白是一种交通运输ATPase，为DNA易位机制以及4型菌毛的组装提供动力。因此，PilF介导了嗜热链球菌的自然转化性。PilF包含一个C端ATPase域和三个N端域，这些域与所谓的一般分泌途径II（GSPII）域具有部分同源性。这三个GSPII域（GSPII-A，GSPII-B和GSPII-C）对于菌毛组装和抽动运动至关重要。它们与霍乱弧菌的ATPase MshE的N末端结构域表现出不同程度的序列同源性它结合细菌第二信使分子c-di-GMP。NMR实验表明，PilF的GSPII-B结构域还以高亲和力结合c-di-GMP，并在NMR时间尺度上缓慢交换形成1：1的复合物。作为对c-di-GMP与嗜热链球菌PilF的GSPII-B结构域结合进行结构研究的先决条件，我们在此处介绍GSPII-B的apo和c-di-GMP结合状态的NMR共振分配。此外，我们使用化学位移扰动数据绘制了c-di-GMP在GSPII-B结构域上的结合位点，并根据以下数据比较了载脂蛋白和GSPII-B结构域的c-di-GMP结合状态的动力学。 { ¹ H}，¹⁵ N-hetNOE数据。