A method to monitor the recruitment of purified LRRK2 kinase onto planar lipid bilayers containing lipid-anchored Rab10 protein using Total Internal Reflection Fluorescence (TIRF) Microscopy.
In this manuscript, in a near-complete pathway from initial membrane recruitment to LC3 lipidation reaction, we show how a three-step targeting mechanism of the ATG12-ATG5-ATG16L1 machinery ensures a high level of regulatory control on autophagy.