Techniques

• Synthesis of inhibitors
• Lead structure-driven, mechanism-based, de novo
• Prodrugs
• PROTACs
• chemical probes


• In vitro assay development
• Fluorescence, fluorescence polarization, time-resolved fluorescence, Alphascreen^R, FRET, luciferase, antibody-based assays, coupled assays
• Synthesis and application of small molecule deacetylase substrates


• Expression and purification of recombinant enzymes; nuclear extracts
• currently: hSIRT1, 2 and 3, PCAF; HDACs from extracts


• Application of assay techniques to in vivo material (hit validation)
  
  • Cell culture, blood, tissue samples
  • Protein acetylation, methylation; enzymatic activity (acetylation)
  • Cellular Thermal Shift Assays (CETSA)
  
  
• In vitro screening (approx. 7000 compounds from external suppliers)


• In silico screening (collaboration with Prof. Dr. Wolfgang Sippl, Halle)

Targets (Inhibitor synthesis and screening)

• Zinc-dependent histone deacetylases (HDACs)
• Recent focus on subtype selectivity (esp. HDAC6, HDAC8, HDAC10, schistosomal HDACs)


• NAD⁺-dependent histone deacetylases (Sirtuins)
• Inhibitors based on focussed library and virtual screening
• Selectivity studies with other NAD⁺-dependent enzymes


• Histone acetyltransferases


• Histone arginine and lysine methyltransferases
• In-silico screening of large databases (Wolfgang Sippl, Halle)
• Functional activity in reporter gene assays (R. Schüle, Freiburg)
• Selectivity testing


• Histone demethylases
• FAD-dependent lysine-specific demethylase 1 (LSD1)
• Iron(II)- and α-ketoglutarate-dependent JumonjiC histone demethylases


• Methyllysine reader proteins
• Inhibitor screening and synthesis
• binding assays


• Early ADME tests (serum protein binding) based on fluorescence polarization


• Numerous collaborations for cellular/animal activity (oncology, neuroscience)

Teaching

• Group seminars
• Lecture series "Assay Development"