PGLD FRAGMENT BASED SCREENING

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 PglD是空肠弯曲杆菌N-连接糖蛋白生物合成途径中的一个重要酶。空肠弯曲菌是人类胃肠炎的主要原因，也是周围神经病格林-巴利综合征之前最常见的感染。已知在缺乏pglD的突变细菌中，空肠弯曲菌特异的蛋白质糖基化被破坏，并且这些细菌感染宿主的能力受到损害。在我们的实验室里，我们启动了一个项目，通过基于片段的筛选来开发一种有效的PglD抑制剂。我们已经确定了11个与PglD结合的小分子(MW&lt；300Da)，并试图通过X射线结晶学来确定它们的结合模式。基本的结构信息将指导开发更紧密的PglD粘结剂，这些粘结剂将在我们的实验室合成和评估。预计这种方法将产生一种特定的、纳米分子的PglD抑制剂。有了这样的工具，我们将能够在体内控制N-糖基化，从而解开它的生物学作用。这种化合物也将具有治疗价值。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. PglD is an essential enzyme in the N-linked glycoprotein biosynthetic pathway in the pathogenic bacteria Campylobacter jejuni. C. jejuni is the primary cause of gastroenteritis in humans and is the most frequent infection to precede the peripheral neuropathy Guillain-Barr¿ syndrome. It is known that the C. jejuni-specific protein glycosylation is disrupted in mutant bacteria lacking pglD, and that these bacteria are compromised in their ability to infect the host organism. In our lab we have initiated a project to develop an effective inhibitor of PglD by fragment-based screening. We have identified eleven small molecules (MW < 300 Da, "fragments") which bind to PglD, and we seek to determine their binding mode by X-ray crystallography. The essential structural information will guide the development of tighter PglD binders which will be synthesized and evaluated in our laboratory. It is anticipated that this approach will result in a specific, nanomolar inhibitor of PglD. Armed with such a tool, we will be able to control N-glycosylation in vivo, thereby unraveling its biological roles. Such a compound will also be of therapeutic value.