al., 2012). Because no association was recorded using either the LPHN3-HR-GAINGPS construct or FLRT3-FN3 domains, we conclude that LPHN3-OLF and FLRT3-LRR represent the complete binding cassette of these proteins. ITC experiments in the presence of 1 mM EDTA or 2 mM Ca2+ showed that the binding affinity remains unchanged (data not shown), indicating that the interaction doesn’t require Ca2+ at the interface, as is the case of neuroligin/neurexin interaction (Fabrichny et al., 2007). The interaction data were interpreted by using the single site model fitting indicating that, in our conditions, the tendency of FLRT3-LRR to dimerize (see below) was abolished by the LPHN3-OLF/FLRT3-LRR association.
