Combination of human acetylcholinesterase and serum albumin sensing surfaces as highly informative analytical tool for inhibitor screening

Fabini, Edoardo; Tramarin, Anna; Bartolini, Manuela

doi:10.1016/j.jpba.2018.03.060

Cited by 12 publications

(7 citation statements)

References 38 publications

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“…They reasoned that the molecules bring in conformational changes upon binding to AChE. Also, immobilization of recombinant human AChE on SPR chip was studied with four known ligands and compared to human serum albumin thereby correlated to the ADME data [22]. Additionally, to determine the kinetic affinity precisely we have used the competitive binding mode of experiments.…”

Section: Discussionmentioning

confidence: 99%

“…There are very limited reports available regarding the screening of AChEI’s by using SPR. Recently, detection of AChEI has been reported with two inhibitors neostigmine and eserine [21] and the affinity of few drug molecules with AChE [22] by using SPR. However, screening and kinetic analysis followed by structure studies of different molecules using SPR for inhibition of AChE can be a fast and rapid alternative to identify potential drug molecules [23–26].…”

Section: Introductionmentioning

confidence: 99%

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Comparative biophysical characterization: A screening tool for acetylcholinesterase inhibitors

Patil

Sistla

et al. 2019

PLoS ONE

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Among neurodegenerative diseases, Alzheimer’s disease (AD) is one of the most grievous disease. The oldest cholinergic hypothesis is used to elevate the level of cognitive impairment and acetylcholinesterase (AChE) comprises the major targeted enzyme in AD. Thus, acetylcholinesterase inhibitors (AChEI) constitutes the essential remedy for the treatment of AD. The study aims to evaluate the interactions between natural molecules and AChE by Surface Plasmon Resonance (SPR). The molecules like alkaloids, polyphenols and substrates of AChE have been considered for the study with a major emphasis on affinity and kinetics. To better understand the activity of small molecules, the investigation is supported by both experimental and theoretical approach such as fluorescence, Circular Dichroism (CD) and molecular docking studies. Amongst the screened ones tannic acid showed promising results compared with others. The methodology followed here have highlighted many molecules with a higher affinity towards AChE and these findings may take lead molecules generated in preclinical studies to treat neurodegenerative diseases. Additionally, we suggest a unique signature for the heterogeneous analyte model using competitive experiments for analyzing simultanous interactions of both the analytes.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Comparative biophysical characterization: A screening tool for acetylcholinesterase inhibitors

Patil

Sistla

et al. 2019

PLoS ONE

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“…Because the motions of amino acid side chains and/or of secondary structure elements within proteins occur at a much slower rate than free diffusion [50], the inhibitor passage through the bottleneck likely constitutes the rate limiting step of the GAL-TcAChE interaction. Recently, employing a surface plasmon resonance (SPR)-based assay, Fabini et al have determined the constant rates for the binding to the human AChE of several inhibitors [51]. The k on and k off for GAL were evaluated to be 9.5 × 10 4 M −1 s −1 , and 2.5 × 10 −2 s −1 respectively, corresponding to a residence time of 40.3 s and to a K i of 0.26 µM.…”

Section: Discussionmentioning

confidence: 99%

Kinetic Modeling of Time-Dependent Enzyme Inhibition by Pre-Steady-State Analysis of Progress Curves: The Case Study of the Anti-Alzheimer’s Drug Galantamine

Lamba

Pesaresi

2022

IJMS

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The Michaelis–Menten model of enzyme kinetic assumes the free ligand approximation, the steady-state approximation and the rapid equilibrium approximation. Analytical methods to model slow-binding inhibitors by the analysis of initial velocities have been developed but, due to their inherent complexity, they are seldom employed. In order to circumvent the complications that arise from the violation of the rapid equilibrium assumption, inhibition is commonly evaluated by pre-incubating the enzyme and the inhibitors so that, even for slow inhibitors, the binding equilibrium is established before the reaction is started. Here, we show that for long drug-target residence time inhibitors, the conventional analysis of initial velocities by the linear regression of double-reciprocal plots fails to provide a correct description of the inhibition mechanism. As a case study, the inhibition of acetylcholinesterase by galantamine, a drug approved for the symptomatic treatment of Alzheimer’s disease, is reported. For over 50 years, analysis based on the conventional steady-state model has overlooked the time-dependent nature of galantamine inhibition, leading to an erroneous assessment of the drug potency and, hence, to discrepancies between biochemical data and the pharmacological evidence. Re-examination of acetylcholinesterase inhibition by pre-steady state analysis of the reaction progress curves showed that the potency of galantamine has indeed been underestimated by a factor of ~100.

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“…Acetylcholinesterase (AChE, E.C. 3.1.1.7) is an essential enzyme for the cleavage of acetylcholine in the nervous system and one of the most used enzymes in biosensors for pesticides (e.g., organophosphates and carbamates) and pharmaceuticals (e.g., a drug for Alzheimer’s disease treatment, donepezil) inhibiting cholinesterase [ 4 , 5 ].…”

Section: Introductionmentioning

confidence: 99%

“…The screening of the kinetic parameters of novel drugs is also a common use of AChE as a biorecognition element [ 5 ]. For the construction of biosensors, it is beneficial to use AChE from Electrophorus electricus in preliminary studies, due to its high similarity to human AChE [ 6 ].…”

Section: Introductionmentioning

confidence: 99%

Open Meter Duo: Low-Cost Instrument for Fluorimetric Determination of Cholinesterase Activity

Keresteš,

Mozo,

Pohanka

2024

Sensors

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Environmental screening is essential due to the increased occurrence of harmful substances in the environment. Open Meter Duo (OMD) is an open-source field photo/fluorimeter that uses an RGB diode that imitates a color according to the selected wavelength and uses a UV LED from the security kit diode as an excitation light source. The prepared PCB shield with a 3D-printed aperture was connected to Arduino UNO R4 WiFi. This system was used for the fluorescent detection of cholinesterase activity with the indoxyl acetate method. Carbofuran—a toxic pesticide—and donepezil—a drug used to treat Alzheimer’s disease—were tested as model inhibitors of cholinesterase activity. The limit of detection of indoxyl acetate was 11.6 μM, and the IC50 values of the inhibitors were evaluated. This system is optimized for wireless use in field analysis with added cloud support and power source. The time of analysis was 5 min for the fluorimetric assay and 20 min for the optional photometric assay. The time of field operation was approximately 4 h of continuous measurement. This system is ready to be used as a cheap and easy control platform for portable use in drug control and point-of-care testing.

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Combination of human acetylcholinesterase and serum albumin sensing surfaces as highly informative analytical tool for inhibitor screening

Cited by 12 publications

References 38 publications

Comparative biophysical characterization: A screening tool for acetylcholinesterase inhibitors

Comparative biophysical characterization: A screening tool for acetylcholinesterase inhibitors

Kinetic Modeling of Time-Dependent Enzyme Inhibition by Pre-Steady-State Analysis of Progress Curves: The Case Study of the Anti-Alzheimer’s Drug Galantamine

Open Meter Duo: Low-Cost Instrument for Fluorimetric Determination of Cholinesterase Activity

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