Coumarin-based turn-on fluorescence probe with a large Stokes shift for detection of endogenous neutrophil elastase in live cells and zebrafish

Sun, Qi; Li, Xiang; Guo, Yun; Qiu, Yuan; Luo, Xiaogang; Liu, Genyan; Han, Yunfeng

doi:10.1016/j.saa.2022.121563

Cited by 7 publications

(3 citation statements)

References 48 publications

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“…It has been reported that pentafluoropropanamide was a specific recognition moiety for neutrophil elastase. − Here, to further elucidate the rationality of adopting heptafluorobutylamide in the chymotrypsin-targeted probe design, we compared the binding model of different TMBIN-based probes. As shown in Figure S7, when the recognition unit of TMBIHF is substituted with trifluoromethyl or pentafluoroethyl, the distances between the catalytic residue and amide bonds of these probes were significantly increased, indicating that heptafluorobutylamide is an appropriate recognition moiety for chymotrypsin.…”

Section: Resultsmentioning

confidence: 99%

De Novo Design of a Highly Selective Nonpeptide Fluorogenic Probe for Chymotrypsin Activity Sensing in a Living System

et al. 2022

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Chymotrypsin, an extensively known proteolytic enzyme, plays a substantial role in maintaining physiological functions, including protein digestion, immune response, and tissue repair. To date, intense attention has been focused on the invention of efficient and sensitive chemical tools for chymotrypsin activity measurement. Among them, the “nonpeptide”-based chymotrypsin probe design strategy utilizing the esterase activity of chymotrypsin has been well-developed due to its low cost and high atom-economy feature. However, the ester-bond-based nature of these probes make them possibly vulnerable to esterases and active chemicals. These defects strictly restricted the application of the previously reported probes, especially for imaging in living systems. Therefore, to acquire fluorogenic probes with sufficient stability and specificity for chymotrypsin sensing in a complicated biological environment, a more stable skeleton for nonpeptide-based chymotrypsin probe construction is urgently needed. Herein, a novel nonpeptide-based fluorogenic probe for specific chymotrypsin activity sensing was designed and synthesized by the substitution of an ester-based linker with a heptafluorobutylamide moiety. The acquired probe, named TMBIHF, showed high selectivity toward various enzymes and reactive chemicals, while it retained high sensitivity and catalytic efficiency toward chymotrypsin. Moreover, TMBIHF was successfully applied for monitoring chymotrypsin activity and pancreas development in live zebrafish, specific sensing of exogenous and endogenous chymotrypsin in nude mice, and visualizing chymotrypsin-like activity-dependent cellular apoptosis, thus providing an alternative and reliable way for chymotrypsin-targeted biosensor or prodrug construction.

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Section: Resultsmentioning

confidence: 99%

De Novo Design of a Highly Selective Nonpeptide Fluorogenic Probe for Chymotrypsin Activity Sensing in a Living System

et al. 2022

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“…Elastases like HNE have been implicated in the destructive degradation of elastin, collagen, proteoglycan and other connective tissue proteins − and are typically found at elevated levels in chronic wounds, which contributes to a myriad of chronic illnesses. , Porcine Pancreatic Elastase (PPE) is often used to study elastase activity for human elastases, including human leukocyte elastase. Both HNE and PPE share 43% sequence homology and have a proline-binding domain with a common recognition motif that is sensitive to electrophilic ketones. , This is regularly accomplished through the aid of fluorescent probes, wherein a fluorophore such as 7-amino-4-methylcoumarin (AMC) is attached to the tripeptide binding domain through a scissile bond and conjugation of AMC via the amino group to the tripeptide results in fluorescent quenching . The enzymatic cleavage of the scissile bond by the elastase gives rise to an increase in the fluorescence signal due to free AMC in solution, which can be used to quantify the enzymatic activity of the elastase.…”

Section: Introductionmentioning

confidence: 99%

“…Both HNE and PPE share 43% sequence homology 23 and have a proline-binding domain with a common recognition motif that is sensitive to electrophilic ketones. 20 , 24 This is regularly accomplished through the aid of fluorescent probes, wherein a fluorophore such as 7-amino-4-methylcoumarin (AMC) is attached to the tripeptide binding domain through a scissile bond and conjugation of AMC via the amino group to the tripeptide results in fluorescent quenching. 25 The enzymatic cleavage of the scissile bond by the elastase gives rise to an increase in the fluorescence signal due to free AMC in solution, which can be used to quantify the enzymatic activity of the elastase.…”

Section: Introductionmentioning

confidence: 99%

Assessment of Cellulose Nanofiber-Based Elastase Biosensors to Inflammatory Disease as a Function of Spacer Length and Fluorescence Response

Easson,

Jordan,

Edwards

et al. 2024

ACS Appl. Bio Mater.

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Inflammatory disease biomarker detection has become a high priority in point-of-care diagnostic research in relation to chronic wounds, with a variety of sensor-based designs becoming available. Herein, two primary aspects of biosensor design are examined: (1) assessment of a cellulose nanofiber (CNF) matrix derived from cotton ginning byproducts as a sensor transducer surface; and (2) assessment of the relation of spacer length and morphology between the CNF cellulose backbone and peptide fluorophore as a function of sensor activity for porcine pancreatic and human neutrophil elastases. X-ray crystallography, specific surface area, and pore size analyses confirmed the suitability of CNF as a matrix for wound care diagnostics. Based upon the normalized degree of substitution, a pegylated-linker connecting CNF transducer substrate to peptide fluorophore showed the greatest fluorescence response, compared to short-and long-chain alkylated linkers.

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Phenoxazine-based red-emitting fluorescence probe for specific detection and imaging of endogenous neutrophil elastase in vitro and in vivo

zhao

Qiu

et al. 2023

Dyes and Pigments

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Coumarin-based turn-on fluorescence probe with a large Stokes shift for detection of endogenous neutrophil elastase in live cells and zebrafish

Cited by 7 publications

References 48 publications

De Novo Design of a Highly Selective Nonpeptide Fluorogenic Probe for Chymotrypsin Activity Sensing in a Living System

De Novo Design of a Highly Selective Nonpeptide Fluorogenic Probe for Chymotrypsin Activity Sensing in a Living System

Assessment of Cellulose Nanofiber-Based Elastase Biosensors to Inflammatory Disease as a Function of Spacer Length and Fluorescence Response

Phenoxazine-based red-emitting fluorescence probe for specific detection and imaging of endogenous neutrophil elastase in vitro and in vivo

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