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Carbonate-Based Fluorescent Chemical Tool for Uncovering Carboxylesterase 1 (CES1) Activity Variations in Live Cells.

Anchal SinghMingze GaoCarolyn J KarnsTaylor P SpidleMichael W Beck
Published in: Chembiochem : a European journal of chemical biology (2022)
Carboxylesterase 1 (CES1) plays a key role in the metabolism of endogenous biomolecules and xenobiotics including a variety of pharmaceuticals. Despite the established importance of CES1 in drug metabolism, methods to study factors that can vary CES1 activity are limited with only a few suitable for use in live cells. Herein, we report the development of FCP1, a new CES1 specific fluorescent probe with a unique carbonate substrate constructed from commercially available reagents. We show that FCP-1 can specifically report on endogenous CES1 activity with a robust fluorescence response in live HepG2 cells through studies with inhibitors and genetic knockdowns. Subsequently, we deployed FCP-1 to develop a live cell fluorescence microscopy-based approach to identify activity differences between CES1 isoforms. To the best of our knowledge, this is the first application of a fluorescent probe to measure the activity of CES1 sequence variants in live cells.
Keyphrases
  • fluorescent probe
  • living cells
  • induced apoptosis
  • cell cycle arrest
  • single molecule
  • healthcare
  • oxidative stress
  • cell death
  • wastewater treatment
  • high throughput
  • dna methylation
  • genome wide
  • pi k akt