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基于蛋白磷酸酶和人工神经网络的 LR 和 YR 微囊藻毒素的高灵敏检测和区分。

Highly sensitive detection and discrimination of LR and YR microcystins based on protein phosphatases and an artificial neural network.

机构信息

Laboratoire IMAGES EA 4218, University of Perpignan Via Domitia, Perpignan, France.

出版信息

Anal Bioanal Chem. 2012 Aug;404(3):711-20. doi: 10.1007/s00216-012-6092-6. Epub 2012 May 31.

Abstract

The inhibition characteristics of three different protein phosphatases by three microcystin (MC) variants--LR, YR, and RR--were studied. The corresponding K (I) for each enzyme-MC couple was calculated. The toxicity of MC varies in the following order: MC-LR > MC-YR > MC-RR. The sensitivity of the enzymes increased in the following order: mutant PP2A < mutant PP1 < natural PP2A. The best limit of detection obtained was 21.2 pM MC-LR using the most sensible enzyme. Methanol, ethanol, and acetonitrile up to 2 % (v/v) may be used in inhibition measurements. An artificial neural network (ANN) was used to discriminate two MC variants--LR and YR--using the differences in inhibition percentages measured with mutant PP1 and natural PP2A. The ANN is able to analyze mixtures with concentrations ranging from 8 to 98 pM MC-LR and 31 to 373 pM MC-YR.

摘要

研究了三种不同的微囊藻毒素(MC)变体-LR、YR 和 RR-对三种不同蛋白磷酸酶的抑制特性。计算了每种酶-MC 偶联物的相应 K(I)。MC 的毒性按以下顺序排列:MC-LR > MC-YR > MC-RR。酶的灵敏度按以下顺序增加:突变型 PP2A < 突变型 PP1 < 天然 PP2A。使用最敏感的酶,获得的最佳检测极限为 21.2 pM MC-LR。甲醇、乙醇和乙腈最高可达 2%(v/v)可用于抑制测量。人工神经网络(ANN)用于使用突变型 PP1 和天然 PP2A 测量的抑制百分比差异来区分两种 MC 变体-LR 和 YR。ANN 能够分析浓度范围为 8 到 98 pM MC-LR 和 31 到 373 pM MC-YR 的混合物。

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