Affiliation:
1. Molecular Sensors and Therapeutics Research Laboratory Department of Chemistry, School of Natural Sciences Shiv Nadar Institute of Eminence (SNIoE) Deemed to be University Delhi NCR, Greater Noida Uttar Pradesh 201314 India
2. Molecular Simulation Research Laboratory Department of Chemistry, School of Natural Sciences Shiv Nadar Institute of Eminence (SNIoE) Deemed to be University Delhi NCR, Greater Noida Uttar Pradesh 201314 India
Abstract
AbstractEsterases enzymes regulate the body's homeostasis by catalyzing the hydrolysis of various esters. These are also involved in protein metabolism, detoxification, and signal transmission. Most importantly, esterase plays a significant role in cell viability and cytotoxicity assays. Hence, developing an efficient chemical probe is essential for monitoring the esterase activity. Several fluorescent probes for esterase have also been reported targeting cytosol and lysosomes. However, the ability to create efficient probes is constrained due to a lack of understanding of the esterase‘s active site for hydrolyzing the substrate. In addition, the fluorescent turn‐on may limit efficient monitoring. Herein, we have developed a unique fluorescent probe, PM‐OAc, to monitor mitochondrial esterase enzyme activity ratiometrically. This probe exhibited a bathochromic wavelength shift with esterase enzyme in alkaline pH (pH∼8.0) due to an intramolecular charge transfer (ICT) process. The phenomenon is well supported by TD‐DFT calculation. Moreover, the substrate (PM‐OAc) binding at the active site of esterase and its catalytic mechanism to hydrolyze the ester bond are elucidated by molecular dynamics (MD) simulation and QM/MM (Quantum mechanics/molecular mechanics) calculations, respectively. Fluorescent image‐based analysis of the cellular environment reveals that our probe can distinguish between live and dead cells based on esterase enzyme activity.
Funder
Science and Engineering Research Board
Shiv Nadar University
Subject
General Chemistry,Catalysis,Organic Chemistry
Cited by
4 articles.
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