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

Prosjektet er et samarbeid mellom Oslo Universitetsykehus, EMBL Outstation Grenoble og ESRF, Grenoble. Målet er å gjennomføre en validering og utprøving av en automatisert plattform for screening av potensielle hemmere mot krystaller av enzymer, og deretter gjennomføre strukturundersøkelser ved hjelp av synkrotronstråling ved ESRF. Vi arbeider med enzymet Ogg1, og hemmere for Ogg1 har et mulig potensiale som adjuvans/hjelpestoff i kreftterapi. Så langt i prosjektet har vi identifisert flere nye ligander for Ogg1, og samlet strukturdata for flere komplekser ved hjelp av den semi-automatiserte plattformen ved EMBL i Grenoble. Videre har vi studert binding mellom enzym og disse ligandene ved hjelp av ulike biofysikalske metoder. Vi arbeider nå med å studere effekt av disse stoffene på celler.

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.