Esterase-Activated Two-Fluorophore System for Ratiometric Sensing of Biological Zinc(II)
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Abstract
Intracellular ester hydrolysis by cytosolic esterases is a common strategy used to trap fluorescent sensors within the cell. We have prepared analogues of Zinpyr-1 (ZP1), an intensity-based fluorescent sensor for Zn2+, that are linked via an amido-ester or diester moiety to a calibrating fluorophore, coumarin 343. These compounds, designated Coumazin-1 and -2, are nonpolar and are quenched by intramolecular interactions between the two fluorophores. Esterase-catalyzed hydrolysis generates a Zn2+-sensitive ZP1-like fluorophore and a Zn2+-insensitive coumarin as a calibrating fluorophore. Upon excitation of the fluorophores, coumarin 343 emission relays information concerning sensor concentration whereas ZP1 emission indicates the relative concentration of Zn2+-bound sensor. This approach enables intracellular monitoring of total sensor concentration and provides a ratiometric system for sensing biological zinc ion.
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