scholarly journals Internally quenched fluorogenic probe provides selective and rapid detection of cathepsin L activity

2020 ◽  
Author(s):  
Kelton A Schleyer ◽  
Ben Fetrow ◽  
Peter Zannes Fatland ◽  
Jun Liu ◽  
Maya Chaaban ◽  
...  
Keyword(s):  
Binding Sites ◽  
Rapid Detection ◽  
Chemical Space ◽  
Cathepsin L ◽  
Turn On ◽  
Disease States ◽  
Fluorogenic Probe ◽  
Quenching Mechanisms ◽  
Ctl Activity ◽  
Enzyme Selectivity

Cathepsin L (CTL) is a cysteine protease that demonstrates upregulated activity and/or altered trafficking during disease states such as cancer. The overlapping substrate specificity of cathepsin family members makes selective detection of activity from a single cathepsin difficult, and CTL activity is particularly difficult to parse from its close homologue CTV and the ubiquitous CTB. Despite this, screening campaigns have explored the extended chemical space in the cathepsin binding sites and identified unique substrate structures that offer selectivity for one enzyme over others. In this vein, we present CTLAP, a fluorogenic probe that is rapidly activated by CTL and displays good selectivity over CTB and CTV, the closest competing analytes for CTL activity probes. CTLAP exhibits intrinsically low background fluorescence, which we attribute to possible self-quenching mechanisms. CTLAP demonstrates markedly higher turn-on ratios (24-fold) and moderately improved enzyme selectivity compared to Z-FR-AMC (10-fold turn-on ratio), a commercially available CTL-selective probe commonly used to detect CTL activity in mixed samples. Optimum selectivity for CTL is achieved within 10 min of incubation with the enzyme, suggesting that CTLAP is amenable for rapid detection of CTL, even in the presence of competing cathepsins.

scholarly journals Intrinsically quenched fluorogenic probe provides selective and rapid detection of cathepsin L activity

ChemMedChem ◽  
2020 ◽  
Author(s):  
Kelton A. Schleyer ◽  
Ben Fetrow ◽  
Peter Zannes Fatland ◽  
Jun Liu ◽  
Maya Chaaban ◽  
...  
Keyword(s):  
Rapid Detection ◽  
Cathepsin L ◽  
Fluorogenic Probe

Abstract 328: Selective fluorogenic probe for rapid detection of cathepsin L activity

2021 ◽  
Author(s):  
Kelton A. Schleyer ◽  
Ben Fetrow ◽  
Peter Fatland ◽  
Jun Liu ◽  
Maya Chaaban ◽  
...  
Keyword(s):  
Rapid Detection ◽  
Cathepsin L ◽  
Fluorogenic Probe

A turn-on fluorescent sensing strategy for rapid detection of Flumequine in water environment using covalent-coordination functionalized MOFs

CrystEngComm ◽  
2021 ◽  
Author(s):  
Shengnan Li ◽  
Ying Li ◽  
Bing Yan
Keyword(s):  
Rapid Detection ◽  
Water Environment ◽  
Fluorescent Sensing ◽  
Turn On ◽  
High Output

With the high output and large use of antibiotics in the process of aquaculture, the pollution caused by antibiotics in water environment is becoming a thorny problem, and its ecological...

scholarly journals A Bioluminescent Sensor for Rapid Detection of PPEP-1, a Clostridioides difficile Biomarker

Sensors ◽  
2021 ◽  
Vol 21 (22) ◽  
pp. 7485
Author(s):  
Kevin K. Ng ◽  
Zachary E. Reinert ◽  
Jeroen Corver ◽  
Danica Resurreccion ◽  
Paul J. Hensbergen ◽  
...  
Keyword(s):  
Ambient Temperature ◽  
Rapid Detection ◽  
Point Of Care ◽  
Delayed Diagnosis ◽  
Digital Camera ◽  
Proteolytic Cleavage ◽  
Fecal Material ◽  
Turn On ◽  
Clostridioides Difficile ◽  
Specific Protease

Current assays for Clostridioides difficile in nonhospital settings are outsourced and time-intensive, resulting in both delayed diagnosis and quarantining of infected individuals. We designed a more rapid point-of-care assay featuring a “turn-on” bioluminescent readout of a C. difficile-specific protease, PPEP-1. NanoLuc, a bright and stable luciferase, was “caged” with a PPEP-1-responsive peptide tail that inhibited luminescence. Upon proteolytic cleavage, the peptide was released and NanoLuc activity was restored, providing a visible readout. The bioluminescent sensor detected PPEP-1 concentrations as low as 10 nM. Sensor uncaging was achieved within minutes, and signal was captured using a digital camera. Importantly, the sensor was also functional at ambient temperature and compatible with fecal material, suggesting that it can be readily deployed in a variety of settings.

scholarly journals A New Fluorescence Probe for Rapid Detection of Amyloid

2018 ◽  
Author(s):  
Nidhi Gour ◽  
Vivek Shinh Kshtriya ◽  
Bharti Koshti ◽  
Shradhey Gupta ◽  
khashti Ballabh Joshi ◽  
...  
Keyword(s):  
Rapid Detection ◽  
Control Experiment ◽  
Fluorescence Probe ◽  
Photophysical Properties ◽  
Cost Effective ◽  
The Novel ◽  
High Quantum Yield ◽  
Turn On ◽  
Heat Treated ◽  
Amyloidogenic Peptides

<p>We report efficient synthesis and photophysical properties of a novel fluorescent pyridothiazole based construct by a facile methodology and its application as a sensor for rapid, selective as well as sensitive detection of amyloid fibres. The novel compound has exceptionally high quantum yield and sensitivity. The fluorescence of pyridothiazole construct is completely quenched (Turn-OFF) upon addition of cupric ions and the fluorescence is recovered on addition of amyloidogenic peptides like A<i>β</i><sub>42 </sub>(Turn-ON). The control experiment with native and heat treated BSA suggest that the new probe binds to beeta/<i>β</i> sheets present in amyloid. The new probe offers direct visualization of fluorescence ‘Turn-OFF’ ‘Turn-ON’ under long UV without the use of sophisticated instrumentation. Hence, a very simple, facile and cost-effective methodology for the rapid detection of amyloid is presented and it may in all possibility pave the way for the design of a better probe for sensing amyloids than conventional ThT.</p>

scholarly journals Hybrid Dynamic Pharmacophore Models as Effective Tools to Identify Novel Chemotypes for Anti-TB Inhibitor Design: A Case Study With Mtb-DapB

2020 ◽  
Vol 8 ◽  
Author(s):  
Chinmayee Choudhury ◽  
Anshu Bhardwaj
Keyword(s):  
Binding Site ◽  
Binding Sites ◽  
Chemical Space ◽  
Pharmacophore Model ◽  
Model Systems ◽  
Receptor Interaction ◽  
Hybrid Molecules ◽  
Wide Range ◽  
Binding Pockets ◽  
Pharmacophore Models

Antimicrobial resistance (AMR) is one of the most serious global public health threats as it compromises the successful treatment of deadly infectious diseases like tuberculosis. New therapeutics are constantly needed but it takes a long time and is expensive to explore new biochemical space. One way to address this issue is to repurpose the validated targets and identify novel chemotypes that can simultaneously bind to multiple binding pockets of these targets as a new lead generation strategy. This study reports such a strategy, dynamic hybrid pharmacophore model (DHPM), which represents the combined interaction features of different binding pockets contrary to the conventional approaches, where pharmacophore models are generated from single binding sites. We have considered Mtb-DapB, a validated mycobacterial drug target, as our model system to explore the effectiveness of DHPMs to screen novel unexplored compounds. Mtb-DapB has a cofactor binding site (CBS) and an adjacent substrate binding site (SBS). Four different model systems of Mtb-DapB were designed where, either NADPH/NADH occupies CBS in presence/absence of an inhibitor 2, 6-PDC in the adjacent SBS. Two more model systems were designed, where 2, 6-PDC was linked to NADPH and NADH to form hybrid molecules. The six model systems were subjected to 200 ns molecular dynamics simulations and trajectories were analyzed to identify stable ligand-receptor interaction features. Based on these interactions, conventional pharmacophore models (CPM) were generated from the individual binding sites while DHPMs were created from hybrid-molecules occupying both binding sites. A huge library of 1,563,764 publicly available molecules were screened by CPMs and DHPMs. The screened hits obtained from both types of models were compared based on their Hashed binary molecular fingerprints and 4-point pharmacophore fingerprints using Tanimoto, Cosine, Dice and Tversky similarity matrices. Molecules screened by DHPM exhibited significant structural diversity, better binding strength and drug like properties as compared to the compounds screened by CPMs indicating the efficiency of DHPM to explore new chemical space for anti-TB drug discovery. The idea of DHPM can be applied for a wide range of mycobacterial or other pathogen targets to venture into unexplored chemical space.

A novel lysosome-targeted fluorogenic probe based on 5-triazole-quinoline for the rapid detection of hydrogen sulfide in living cells

2018 ◽  
Vol 16 (5) ◽  
pp. 712-716 ◽  
Author(s):  
Yandong Dou ◽  
Xiaoxu Gu ◽  
Shasha Ying ◽  
Shen Zhu ◽  
Shian Yu ◽  
...  
Keyword(s):  
Hydrogen Sulfide ◽  
Rapid Detection ◽  
Living Cells ◽  
The Novel ◽  
Signaling Molecule ◽  
Fluorogenic Probe

A fluorogenic probe based on the novel fluorophore 5-triazole-quinoline was developed for the detection of hydrogen sulfide, an endogenous signaling molecule associated with the development of various diseases.

Ligand-displacement-based two-photon fluorogenic probe for visualizing mercapto biomolecules in live cells, Drosophila brains and zebrafish

The Analyst ◽  
2018 ◽  
Vol 143 (14) ◽  
pp. 3433-3441 ◽  
Author(s):  
Yanfei Zhao ◽  
Yun Ni ◽  
Liulin Wang ◽  
Chenchen Xu ◽  
Chenqi Xin ◽  
...  
Keyword(s):  
Rapid Detection ◽  
Live Cell ◽  
Live Cells ◽  
Two Photon ◽  
Ligand Displacement ◽  
Cell Tissue ◽  
Fluorogenic Probe ◽  
Displacement Based

We report the Fe(iii)-based complex TPFeS which acts as a novel ligand-displacement-based TP fluorogenic probe for the rapid detection of mercapto biomolecules both in vitro and in live cell/tissue/in vivo imaging.

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