Abstract 2045 Survival analysis of PI effector binding reveals multiple affinity states

The recruitment of effector proteins to the plasma membrane (PM) by binding to phosphoinositides (PIs) such as PI(4,5)P2 and PI(4)P is integral to myriad cellular processes, with the membrane association and dissociation rates kon and koff for individual binding domains potentially being a factor in the appropriate regulation of these processes. The purpose of this project was to understand binding kinetics of lipid effectors and how they might relate to cellular function. To achieve this, we employed genetically encoded lipid biosensors tagged with mStayGold, resolvable at single molecule levels and imaged at a temporal resolution of 30 ms using TIRF microscopy.