RRC ID 28400
Author Ward CJ, Beattie KA, Lee EY, Codd GA.
Title Colorimetric protein phosphatase inhibition assay of laboratory strains and natural blooms of cyanobacteria: comparisons with high-performance liquid chromatographic analysis for microcystins.
Journal FEMS Microbiol Lett
Abstract Microcystins are cyclic heptapeptide hepatotoxins commonly produced by bloom-forming genera of cyanobacteria. These toxins are potent and specific inhibitors of protein phosphatases 1 and 2A. We have optimised a rapid, simple and sensitive colorimetric protein phosphatase 1 inhibition assay, utilising the activity of protein phosphatase 1 as expressed in a recombinant strain of Escherichia coli, towards the chromogenic substrate, p-nitrophenyl phosphate. A standard curve for the inhibition of protein phosphatase 1 by microcystin-LR was constructed with an IC50 of about 38 ng ml-1 and a limit of detection of 10-20 ng ml-1. Twenty-three laboratory-grown strains and 25 natural bloom samples of cyanobacteria were analysed by high-performance liquid chromatography for microcystins and by the protein phosphatase 1 inhibition assay. Agreement for the microcystin contents of the samples detected by high-performance liquid chromatography and the protein phosphatase 1 inhibition assay showed good correlation (R2 > 0.93, P < 0.0001). The suitability of the colorimetric protein phosphatase 1 inhibition assay as a screen for cyanobacterial microcystins is discussed.
Volume 153(2)
Pages 465-73
Published 1997-8-15
DOI 10.1016/s0378-1097(97)00290-5
PII S0378-1097(97)00290-5
PMID 9271876
MeSH Chromatography, High Pressure Liquid / methods* Colorimetry / methods* Cyanobacteria / chemistry* Enzyme Inhibitors / pharmacology Escherichia coli / genetics Indicators and Reagents Methanol Microcystins Nitrophenols Organophosphorus Compounds Peptides, Cyclic / analysis* Peptides, Cyclic / pharmacology Phosphoprotein Phosphatases / antagonists & inhibitors* Protein Phosphatase 1 Sensitivity and Specificity Solvents
IF 1.987
Times Cited 120
Algae NIES-42 NIES-103 NIES-104 NIES-595 NIES-298 NIES-102