Rapid screening of monoamine oxidase B inhibitors in natural extracts by capillary electrophoresis after enzymatic reaction at capillary inlet

Hu, Kun; Zhang, Lichun; Li, Xiangtang; Zhao, Shulin

doi:10.1016/j.jchromb.2010.09.030

Cited by 25 publications

(9 citation statements)

References 17 publications

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“…CE is a powerful separation tool with the advantage of high‐efficiency separation, short analysis times and low sample loading , and studies have shown that CE is a versatile platform for enzyme study and inhibitor screening . Chenet et al developed a CE method for the screening of 20 proteasome inhibitors .…”

Section: Introductionmentioning

confidence: 99%

Preparation of a dual‐enzyme co‐immobilized capillary microreactor and simultaneous screening of multiple enzyme inhibitors by capillary electrophoresis

Lin

Zhao

et al. 2013

J of Separation Science

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A CE method based on a dual-enzyme co-immobilized capillary microreactor was developed for the simultaneous screening of multiple enzyme inhibitors. The capillary microreactor was prepared by co-immobilizing adenosine deaminase and xanthine oxidase on the inner wall at the inlet end of the separation capillary. The enzymes were first immobilized on gold nanoparticles, and the functionalized gold nanoparticles were then assembled on the inner wall at the inlet end of the separation capillary treated with polyethyleneimine. With the developed CE method, the substrates and products were baseline separated within 3 min. The activity of the immobilized enzyme can be directly detected by measuring the peak height of the products. A statistical parameter Z' factor was recommended for evaluation of the accuracy of a drug screening system. In the present study, it was calculated to be larger than 0.5, implying a good accuracy. Finally, screening a small compound library containing two known enzyme inhibitors and 20 natural extracts by the proposed method was demonstrated. The known inhibitors were identified, and some natural extracts were found to be positive for two-enzyme inhibition by the present method.

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Section: Introductionmentioning

confidence: 99%

Preparation of a dual‐enzyme co‐immobilized capillary microreactor and simultaneous screening of multiple enzyme inhibitors by capillary electrophoresis

Lin

Zhao

et al. 2013

J of Separation Science

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“…Therefore, studying MAO enzymes and finding new MAO inhibitors as purported neuroprotectants is a subject of current interest in drug discovery 1,[18][19][20][21][22][23][24][25][26] . MAO enzymes are usually monitored by measuring the absorbance and/or fluorescence of the oxidation products generated from key amines (kynuramine, tryptamine, tyramine, etc) [27][28][29][30][31][32][33] . The purpose of this research was to evaluate the oxidation of the neurotoxin MPTP by human MAO isozymes (human MAO-A and B) through the chromatographic analysis of the toxic pyridinium species (MPDP + and MPP + ) generated, and subsequently use this approach as a new tool to search for new MAO inhibitors (MAO-A and B) and eventual protective agents.…”

Section: Spanish National Research Council (Csic) Instituto De Cienmentioning

confidence: 99%

Evaluation of the oxidation of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) to toxic pyridinium cations by monoamine oxidase (MAO) enzymes and its use to search for new MAO inhibitors and protective agents

Herraiz

2011

Journal of Enzyme Inhibition and Medicinal Chemistry

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“…CE‐based enzyme assays are economical and efficient, and they require only small volumes of biochemical reagents, an aspect that is particularly beneficial when enzyme and substrates are very expensive. An in‐capillary enzyme assay denominated as the electrophoretically mediated microanalysis (EMMA) has been proven to be very significant for studying enzyme reactions and for enzyme inhibitor screening . This assay is performed by injecting nanoliter volumes of enzyme and substrate solutions orderly into the capillary.…”

Section: Introductionmentioning

confidence: 99%

Rapid screening of aminopeptidase N inhibitors by capillary electrophoresis with electrophoretically mediated microanalysis

Wang

Cao

et al. 2014

Electrophoresis

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In this work, an electrophoretically mediated microanalysis (EMMA) method with a partial-filling technique was setup to evaluate the inhibitory potency of novel compounds toward aminopeptidase N (APN). It was necessary to optimize the electrophoretic conditions with respect to the kinetic constraints and for attaining high sensitivity. In our setup, a part of the capillary was filled with the incubation buffer for the enzyme reaction, whereas the rest was filled with a suitable BGE for the separation of substrates and products. To monitor the performance of the newly developed method, the kinetic constants (Km and Vmax) for the catalyzed dissociation of L-Leucine-p-nitroanilide in the presence of APN as well as the inhibition constant (IC50 ) of a known competitive inhibitor, that is bestatin, were determined and these results were compared with those obtained by a classical spectrophotometric assay. The developed EMMA method was subsequently applied to the screening of 30 APN inhibitors. Whereas the inhibition potency of these inhibitors (expressed in IC50 values) were significantly underestimated by the EMMA method, the order of the inhibitory potential of these various compounds was found in agreement with the literature.

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Rapid screening of monoamine oxidase B inhibitors in natural extracts by capillary electrophoresis after enzymatic reaction at capillary inlet

Cited by 25 publications

References 17 publications

Preparation of a dual‐enzyme co‐immobilized capillary microreactor and simultaneous screening of multiple enzyme inhibitors by capillary electrophoresis

Preparation of a dual‐enzyme co‐immobilized capillary microreactor and simultaneous screening of multiple enzyme inhibitors by capillary electrophoresis

Evaluation of the oxidation of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) to toxic pyridinium cations by monoamine oxidase (MAO) enzymes and its use to search for new MAO inhibitors and protective agents

Rapid screening of aminopeptidase N inhibitors by capillary electrophoresis with electrophoretically mediated microanalysis

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