Tsao Désirée H H, Sutherland Alan G, Jennings Lee D, Li Yuanhong, Rush Thomas S, Alvarez Juan C, Ding Weidong, Dushin Elizabeth G, Dushin Russell G, Haney Steve A, Kenny Cynthia H, Malakian A Karl, Nilakantan Ramaswamy, Mosyak Lidia
Structural Biology and Computational Chemistry, Wyeth Research, Cambridge, MA 02140, USA.
Bioorg Med Chem. 2006 Dec 1;14(23):7953-61. doi: 10.1016/j.bmc.2006.07.050. Epub 2006 Aug 17.
ZipA is a membrane anchored protein in Escherichia coli that interacts with FtsZ, a homolog of eukaryotic tubulins, forming a septal ring structure that mediates bacterial cell division. Thus, the ZipA/FtsZ protein-protein interaction is a potential target for an antibacterial agent. We report here an NMR-based fragment screening approach which identified several hits that bind to the C-terminal region of ZipA. The screen was performed by 1H-15N HSQC experiments on a library of 825 fragments that are small, lead-like, and highly soluble. Seven hits were identified, and the binding mode of the best one was revealed in the X-ray crystal structure. Similar to the ZipA/FtsZ contacts, the driving force in the binding of the small molecule ligands to ZipA is achieved through hydrophobic interactions. Analogs of this hit were also evaluated by NMR and X-ray crystal structures of these analogs with ZipA were obtained, providing structural information to help guide the medicinal chemistry efforts.
ZipA是大肠杆菌中的一种膜锚定蛋白，它与真核微管蛋白的同源物FtsZ相互作用，形成一个介导细菌细胞分裂的隔膜环结构。因此，ZipA/FtsZ蛋白-蛋白相互作用是抗菌剂的一个潜在靶点。我们在此报告一种基于核磁共振的片段筛选方法，该方法鉴定出了几个与ZipA C末端区域结合的命中片段。筛选是通过对一个包含825个片段的文库进行1H-15N HSQC实验来进行的，这些片段体积小、类似先导化合物且高度可溶。鉴定出了7个命中片段，并通过X射线晶体结构揭示了最佳命中片段的结合模式。与ZipA/FtsZ的接触类似，小分子配体与ZipA结合的驱动力是通过疏水相互作用实现的。还通过核磁共振对该命中片段的类似物进行了评估，并获得了这些类似物与ZipA的X射线晶体结构，为指导药物化学研究提供了结构信息。
