Rational design of
practical probes with excellent specificity
and improved optical properties for a particular enzyme is always
a big challenge. Herein, a practical and highly specific fluorescent
probe for carboxylesterase 1 (CES1) was rationally designed using meso-carboxyl-BODIPY as the basic fluorophore based on the
substrate preference and catalytic properties of CES1. Following molecular
docking-based virtual screening combined with reaction phenotyping-based
experimental screening, we found that MMB (probe 7) exhibited
the optimal combination of sensitivity and specificity toward human
CES1 in contrast to other ester derivatives. Under physiological conditions,
MMB could be readily hydrolyzed by CES1 and release MCB; such biotransformation
brought great changes in the electronic properties at the meso position of the fluorophore and triggered a dramatic
increase in fluorescence emission around 595 nm. Moreover, MMB was
cell membrane permeable and was successfully applied to monitor the
real activities of CES1 in various biological samples including living
cells, tissue slices, organs, and zebrafish. In summary, this study
showed a good example for constructing specific fluorescent probe(s)
for a target enzyme and also provided a practical and sensitive tool
for real-time sensing of CES1 activities in complicated biological
samples. All these findings would strongly facilitate high-throughput
screening of CES1 modulators and the studies on CES1-associated physiological
and pathological processes.
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