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.
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The work was supported by the Romania–France bilateral grant Brancusi (487/2011). O.I. Covaci is a PhD student with a scholarship funded by the Sectoral Operational Programme Human Resources Development 2007–2013 of the Romanian Ministry of Labour, Family and Social Protection through the Financial Agreement POSDRU/6/1.5/S/19.
Published in the special paper collection Instrumental Methods of Analysis (IMA 2011) with guest editors Maria Ochsenkuehn-Petropoulou, Nikos Kallithrakas, and Panagiotis Kefalas.
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Covaci, O.I., Sassolas, A., Alonso, G.A. et al. Highly sensitive detection and discrimination of LR and YR microcystins based on protein phosphatases and an artificial neural network. Anal Bioanal Chem 404, 711–720 (2012). https://doi.org/10.1007/s00216-012-6092-6
- Protein phosphatase
- Artificial neural network
- Influence of organic solvents