Construction and optimization of a family of genetically encoded metabolite sensors by semirational protein engineering.

ABSTRACT: A family of genetically-encoded metabolite sensors has been constructed using bacterial periplasmic binding proteins (PBPs) linearly fused to protein fluorophores. The ligand-induced conformational change in a PBP allosterically regulates the relative distance and orientation of a fluorescence resonance energy transfer (FRET)-compatible protein pair. Ligand binding is transduced into a macroscopic FRET observable, providing a reagent for in vitro and in vivo ligand-measurement and visualization. Sensors with a higher FRET signal change are required to expand the dynamic range and allow visualization of subtle analyte changes under high noise conditions. Various observations suggest that factors other than inter-fluorophore separation contribute to FRET transfer efficiency and the resulting

SUBMITTER: Deuschle K 

PROVIDER: S-EPMC2253473 | biostudies-literature | 2005 Sep

REPOSITORIES: biostudies-literature