Inhibition equivalency factors for microcystin variants in recombinant and wild-type protein phosphatase 1 and 2A assays
Garibo, Diana (Institut de Recerca i Tecnologia Agroalimentàries)
Flores, Cintia (Institut de Diagnosi Ambiental i Estudis de l'Aigua)
Cetó Alsedà, Xavier (Universitat Autònoma de Barcelona. Departament de Química)
Prieto-Simón, Beatriz (Universitat Autònoma de Barcelona. Departament de Química)
Valle Zafra, Manuel del (Universitat Autònoma de Barcelona. Departament de Química)
Caixach, Josep (Institut de Diagnosi Ambiental i Estudis de l'Aigua)
Diogène, Jorge (Institut de Recerca i Tecnologia Agroalimentàries)
Campàs i Homs, Mònica (Institut de Recerca i Tecnologia Agroalimentàries)

Date:	2014
Abstract:	In this work, protein phosphatase inhibition assays (PPIAs) have been used to evaluate the performance of recombinant PP1 and recombinant and wild-type PP2As. The enzymes have been compared using microcystins-LR (MC-LR) as a model cyanotoxin. Whereas PP2A Rec provides a limit of detection (LOD) of 3. 1 μg/L, PP1 Rec and PP2A Wild provide LODs of 0. 6 and 0. 5 μg/L, respectively, lower than the guideline value proposed by the World Health Organization (1 μg/L). The inhibitory potencies of seven MC variants (-LR, -RR, -dmLR, -YR, -LY, -LW and -LF) have been evaluated, resulting on 50 % inhibition coefficient (IC 50 ) values ranging from 1. 4 to 359. 3 μg/L depending on the MC variant and the PP. The PPIAs have been applied to the determination of MC equivalent contents in a natural cyanobacterial bloom and an artificially contaminated sample, with multi-MC profiles. The inhibition equivalency factors (IEFs) have been applied to the individual MC quantifications determined by liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis, and the estimated MC-LR equivalent content has been compared to PPIA results. PPIAs have demonstrated to be applicable as MC screening tools for environmental applications and to protect human and animal health.
Grants:	Ministerio de Ciencia e Innovación 2008-00084-00-00 Ministerio de Ciencia e Innovación BIO2011-26311
Note:	Altres ajuts: ICREA Academia
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Language:	Anglès
Document:	Article ; recerca ; Versió acceptada per publicar
Subject:	Inhibition equivalency factors (IEFs) ; Liquid chromatography-tandem mass spectrometry (LC-MS/MS) ; Microcystins (MCs) ; Multivariate data analysis ; Protein phosphatase 1 (PP1) ; Protein phosphatase 2A (PP2A) ; Protein phosphatase inhibition assay (PPIA) ; SDG 3 - Good Health and Well-being
Published in:	Environmental science and pollution research international, Vol. 21, Issue 18 (September 2014) , p. 10652-10660, ISSN 1614-7499

DOI: 10.1007/s11356-014-3065-7

Postprint 22 p, 444.8 KB

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