Development, implementation and validation of a platform for structure-based automatic ligand screening

The project is a collaboration between Oslo University Hospital, EMBL Outstation Grenble and ESRF, Grenoble. The aim is to test and validate an automated platform for screening of potential enzyme inhibitors using crystals of the target protein, and then analyse these samples by X-ray crystallography using the synchrotron radiation provided by the collaborator ESRF. We have selected human Ogg1 as Our target of interest, and inhibitors of Ogg1 might have a potential as adjuvants in cancer therapy. So far we have identified a series of novel Ogg1 ligands, and collected structural data for several complexes using the semi-automated platform at EMBL, Grenoble. Further, we have characterized the binding between enzyme and ligands using various biophysical methods. We are now in the process of studying the effect of these compounds on cells.

The present application seeks to develop, streamline, validate and build competence in using high-throughput methods for automatic fragment/ligand screening, data collection and crystal structure analysis as tools within a structure based drug design pipe line. The most time consuming and challenging steps in fragment based drug discovery using X-ray diffraction as an analytic tool, are sample preparation/mounting and data processing/analysis. In this application, we will develop and optimize protocols, an d validate an automated setup for crystal handling - the CrystalDirect technology - in combination with various fragment libraries. The project will be carried out in close collaboration with the EMBL High Throughput Crystallization (HTX) Laboratory and t he ESRF Structural Biology (MX) group, which are both located at the ESRF Synchrotron in Grenoble. The PhD candidate will spend ~12 months in the HTX and MX laboratories at ESRF to take part in the development, optimization and validation of the automatic ligand screening, including data collection and analysis. The project will focus on fragment/ligand screening and crystal structure determination of protein-ligand complexes to identify possible inhibitors for human DNA glycosylases. Such compounds have potential applications as adjuvants to sensitize cancer cells for increased efficacy of current cancer therapies. The methods and competence developed in this project will be communicated to users of our Regional Core Facility for Structural Biology and B ioinformatics, and included in our platform portfolio. By connecting this application and transfer of knowledge to our Core Facility, there will be a substantial synergy between this project and future related projects at the national level.