A Novel Approach to the Development of Highly Specific Inhibitors of a Critical Transcription Factor in Prostate Cancer
Final rept. 15 Feb 2007-14 Feb 2009
VIRGINIA UNIV CHARLOTTESVILLE
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Using isothermal titration calorimetry ITC, we have measured the binding affinity of several constructs of ERG to DNA. The results show that there is auto-inhibition of DNA binding by regions outside the conserved Ets domain, as observed for Ets-1. Based on this data, we are focusing our structural efforts on a fragment of ERG which is auto-inhibited. Backbone NMR resonance assignments for this fragment have been completed and sidechain assignments are in progress. Analysis of the backbone chemical shifts shows the expected fold for the Ets domain. Regions outside the Ets domain show some evidence of additional structural elements but will require the complete structure determination for full characterization. We are well positioned now to complete the structure determination of the auto-inhibited form. We have also developed and validated a fluorescence polarization assay for screening effects of compounds which can be used for fragment screening and high throughput screening.
- Medicine and Medical Research