Development of three-colour FRET cascade for force sensing of the putative RIAM-vinculin interaction in fibroblasts.

ABSTRACT: Förster resonance energy transfer (FRET) enables the measurement of molecular interactions and conformational dynamics in biological systems. FRET-cascade, a multistep energy transfer system involving three fluorophores, enables spatial and temporal mapping of molecular interactions. Here, we leveraged FRET-cascade with time-correlated single photon counting fluorescence lifetime imaging microscopy (TCSPC-FLIM) to explore the putative interaction between Rap1-interacting Adaptor Molecule (RIAM) and vinculin in focal adhesions. We developed a novel three-fluorophore FRET-cascade system, validated using purified proteins, spectroscopic analysis, structural modelling, and negative-staining transmission electron microscopy (TEM). Putative RIAM-vinculin interactions were explored in vinculin kn

SUBMITTER: Treacy CA 

PROVIDER: S-EPMC12828031 | biostudies-literature | 2025 Dec

REPOSITORIES: biostudies-literature