Molecular structural studies of protein complexes essential for viral replication and plant nitrogen metabolism

蛋白质之间的相互作用对许多重要的生物过程都是至关重要的。在病毒复制和植物新陈代谢调控这两个不同的领域，我们正在建立一个分子结构基础，以了解这些过程背后的蛋白质-蛋白质和蛋白质-RNA相互作用的复杂网络。为了实现这一目标，我们开发了有效的方法来生产毫克量的中心酶和调节蛋白。一套生化和生物物理方法也被用来测量结合相互作用和酶活性。最后，结晶筛选和X射线结晶学被用来提供关于蛋白质复合体的高分辨率三维信息。这些结构信息为进一步探索特定相互作用在病毒复制和植物代谢中的重要性的诱变实验设计提供了合理的基础。最终，这些信息也为开发针对重要人畜疾病的新型抗病毒药物以及在氮素限制条件下改良植物作物提供了基础。

Protein-protein interactions are critical for many important biological processes. In the two divergent fields of viral replication and the regulation of plant metabolism, we are establishing a molecular structural basis for understanding the complex network of protein-protein and protein-RNA interactions underlying these processes. To achieve this goal, we have developed efficient methods for producing milligram quantities of the central enzymes and regulatory proteins. A suite of biochemical and biophysical methods have also being used to measure binding interactions and enzyme activities. Finally, crystallization screening and X-ray crystallography are used to provide high resolution three-dimensional information on protein complexes. This structural information provides a rational basis for the design of mutagenesis experiments to further probe the importance of specific interactions in both viral replication and plant metabolism. Ultimately, this information also provides the basis for developing novel antiviral therapeutics for important human and animal diseases, as well as for the improvement of plant crops under conditions of nitrogen limitation.