Mechanisms of Protein-Protein Binding: Double-Mutant Cycle Analysis Using the ProteOn XPR36 System

interactions. Schreiber and colleagues are using the multiplemutant cycle method to evaluate cooperativity of residue binding (Reichmann et al. 2005). With this method, the binding of two mutant proteins is measured individually and together; the interaction energy between the two proteins can then be determined by subtracting the free energy difference in the binding of a double-mutant protein from that of the two singlemutant proteins. The data presented in this tech note is in the context of this larger study.