Immobilization of Recombinant Fluorescent Biosensors Permits Imaging of Extracellular Ion Signals.

ABSTRACT: Given the importance of ion gradients and fluxes in biology, monitoring ions locally at the exterior of the plasma membrane of intact cells in a noninvasive manner is highly desirable but challenging. Classical targeting of genetically encoded biosensors at the exterior of cell surfaces would be a suitable approach; however, it often leads to intracellular accumulation of the tools in vesicular structures and adverse modifications, possibly impairing sensor functionality. To tackle these issues, we generated recombinant fluorescent ion biosensors fused to traptavidin (TAv) specifically coupled to a biotinylated AviTag expressed on the outer cell surface of cells. We show that purified chimeras of TAv and pH-Lemon or GEPII 1.0, Förster resonance energy transfer-based pH and K⁺ bi

SUBMITTER: Burgstaller S 

PROVIDER: S-EPMC8630794 | biostudies-literature | 2021 Nov

REPOSITORIES: biostudies-literature