Understanding Docking Complexes of Macromolecules Using HADDOCK: The Synergy between Experimental Data and Computations

This protocol illustrates the modelling of a protein-peptide complex using the synergic combination of in silico analysis and experimental results. To this end, we use the integrative modelling software HADDOCK, which possesses the powerful ability to incorporate experimental data, such as NMR Chemical Shift Perturbations and biochemical protein-peptide interaction data, as restraints to guide the docking process. Based on the modelling results, a rational mutagenesis approach is used to validate the generated models. The experimental results allow to select a final structural model best representing the bona fide protein-peptide complex. The described protocol can also be applied to model protein-protein complexes. There is no size limit for the macromolecular complexes that can be characterized by HADDOCK as long as the 3D structures of the individual components are available. Keywords: HADDOCK, Molecular docking, Integrative Modelling, Biomolecular interactions, Macromolecular complex, HCN channels, TRIP8b, ITC, NMR, Rational mutagenesi