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
|
WOS Category
|
MICROBIOLOGY
|
Resource |
Algae |
NIES-42
NIES-103
NIES-104
NIES-595
NIES-298
NIES-102 |