Johann Wolfgang Goethe University Frankfurt 2 articles published in JoVE Biochemistry NMR-Based Fragment Screening in a Minimum Sample but Maximum Automation Mode Hannes Berg*1, M. A. Wirtz Martin*1, A. Niesteruk1,2, C. Richter1, S. Sreeramulu1, H. Schwalbe1,2 1Institute for Organic Chemistry and Chemical Biology, Center for Biomolecular Magnetic Resonance (BMRZ), Johann Wolfgang Goethe-University Frankfurt, 2German Cancer Consortium (DKTK) and DKFZ Fragment-based screening by NMR is a robust method to rapidly identify small molecule binders to biomacromolecules (DNA, RNA, or proteins). Protocols describing automation-based sample preparation, NMR experiments & acquisition conditions, and analysis workflows are presented. The technique allows for optimal exploitation of both 1H and 19F NMR-active nuclei for detection. Biochemistry Nano-Differential Scanning Fluorimetry for Screening in Fragment-based Lead Discovery Misbha Ud Din Ahmad1, Alexander Fish1, Jeroen Molenaar1, Sridhar Sreeramulu2, Christian Richter2, Nadide Altincekic2, Harald Schwalbe2, Hans Wienk1, Anastassis Perrakis1 1Oncode Institute and Division of Biochemistry, the Netherlands Cancer Institute, 2Institute for Organic Chemistry and Chemical Biology, Center for Biomolecular Magnetic Resonance (BMRZ), Johann Wolfgang Goethe-University Monitoring changes in the melting temperature of a target protein (i.e., thermal shift assay, TSA) is an efficient method for screening fragment libraries of a few hundred compounds. We present a TSA protocol implementing robotics-assisted nano-Differential Scanning Fluorimetry (nano-DSF) for monitoring intrinsic tryptophan fluorescence and light back-scattering for fragment screening.
