Fluorescent Binding Protein Sensors for Detection and Quantification of Biochemicals, Metabolites, and Natural...

Membrane transporters and soluble binding proteins recognize particular nutrients, metabolites, vitamins, or ligands. By modifying genetically engineered single cysteine residues near the active sites of such proteins with extrinsic maleimide fluorophores,the approaches that we report create sensitive fluorescent sensors that detect, quantify, and monitor molecules that are relevant to the biochemistry, physiology, microbiology, and clinical properties of pro- and eukaryotic organisms. This protocol was validated in: J Biol Chem (2022), DOI: 10.1016/j.jbc.2022.101651 Graphical abstract.