Macromolecular interactions in vitro, comparing classical and novel approaches

Biophysical quantification of protein interactions is central to unveil the molecular mechanisms cellular processes. Researchers can choose from a wide panel biophysical methods that quantify in different ways, including both classical and more novel techniques. We report outcome an ARBRE-MOBIEU training school held June 2019 Gif-sur-Yvette, France ( https://mosbio.sciencesconf.org/ ). Twenty European students benefited week’s with theoretical practical sessions six complementary approaches: (1) analytical ultracentrifugation or without fluorescence detector system (AUC-FDS), (2) isothermal titration calorimetry (ITC), (3) size exclusion chromatography coupled multi-angle light scattering (SEC-MALS), (4) bio-layer interferometry (BLI), (5) microscale thermophoresis (MST) and, (6) switchSENSE. They implemented all these on two examples macromolecular nanomolar affinity: first, protein–protein interaction between artificial alphaRep binder, its target protein, also alphaRep; second, protein-DNA DNA repair complex, Ku70/Ku80 (hereafter called Ku), cognate ligand. approaches used analyze systems under study thereby showcase application each The workshop provided improved understanding advantages limitations methods, enabling future choices concerning are most relevant informative for specific kinds sample interaction.