Defining phospho-Rab signalling networks downstream of LRRK2 kinase activation

TBC1D2 is a newly identified protein that interacts with phosphorylated Rab GTPases. Here we have performed a thermodynamic experiment to understand the interaction between this protein and the globular G domain of phosphorylated Rab8A. The Rab was in vitro phosphorylated at Thr72, purified and the interaction with a coiled coil region of TBC1D2 (residues 353-436, with a NusA solubility tag at the N-terminus). The two proteins were dialized together and pRab8A was injected into a cell containing Nus-TBC1D2 (coiled coil region) using an ITC200 calorimeter from Particular Sciences. Data were processed using Origin 7.0 software and the final files in OPj format are provided here, for each titration.