Quantitative FRET Microscopy Reveals a Crucial Role of Cytoskeleton in Promoting PI(4,5)P₂ Confinement.

ABSTRACT: Phosphatidylinositol 4,5-bisphosphate (PI(4,5)P₂) is an essential plasma membrane component involved in several cellular functions, including membrane trafficking and cytoskeleton organization. This function multiplicity is partially achieved through a dynamic spatiotemporal organization of PI(4,5)P₂ within the membrane. Here, we use a Förster resonance energy transfer (FRET) approach to quantitatively assess the extent of PI(4,5)P₂ confinement within the plasma membrane. This methodology relies on the rigorous evaluation of the dependence of absolute FRET efficiencies between pleckstrin homology domains (PH_PLCδ) fused with fluorescent proteins and their average fluorescence intensity at the membrane. PI(4,5)P₂ is found to be significa

SUBMITTER: Sarmento MJ 

PROVIDER: S-EPMC8583820 | biostudies-literature | 2021 Oct

REPOSITORIES: biostudies-literature