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Engineering of a Red Fluorogenic Protein/Merocyanine Complex for Live‐Cell Imaging

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A reengineered human cellular retinol binding protein II (hCRBPII), a 15‐kDa protein belonging to the intracellular lipid binding protein (iLBP) family, generates a highly fluorescent red pigment through the covalent linkage… Click to show full abstract

A reengineered human cellular retinol binding protein II (hCRBPII), a 15‐kDa protein belonging to the intracellular lipid binding protein (iLBP) family, generates a highly fluorescent red pigment through the covalent linkage of a merocyanine aldehyde to an active site lysine residue. The complex exhibits “turn‐on” fluorescence, due to a weakly fluorescent aldehyde that “lights up” with subsequent formation of a strongly fluorescent merocyanine dye within the binding pocket of the protein. Cellular penetration of merocyanine is rapid, and fluorophore maturation is nearly instantaneous. The hCRBPII/merocyanine complex displays high quantum yield, low cytotoxicity, specificity in labeling organelles, and compatibility in both cancer cell lines and yeast cells. The hCRBPII/merocyanine tag is brighter than most common red fluorescent proteins.

Keywords: protein; merocyanine complex; engineering red; merocyanine; red fluorogenic; cell

Journal Title: ChemBioChem
Year Published: 2019

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