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https://hdl.handle.net/2440/14423
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Type: | Journal article |
Title: | Cell-free protein synthesis inhibition assay for the cyanobacterial toxin cylindrospermopsin |
Author: | Froscio, S. Humpage, A. Burcham, P. Falconer, I. |
Citation: | Environmental Toxicology, 2001; 16(5):408-412 |
Publisher: | John Wiley & Sons Inc |
Issue Date: | 2001 |
ISSN: | 1520-4081 1522-7278 |
Abstract: | The cyanobacterial toxin cylindrospermopsin (CYN) is known to be a potent inhibitor of protein synthesis. This paper describes the use of a rabbit reticulocyte lysate translation system as a protein synthesis inhibition assay for CYN. A dose response curve for protein synthesis inhibition by CYN was constructed and was modeled to a sigmoidal dose response curve with variable slope (R2 = 0.98). In this assay, CYN has an IC50 of 120 nM [95% confidence limits (Cl) = 111-130 nM] with a detection limit in the region of 50 nM in the assay solution. Application of the assay allows quantification of toxin samples within the range 0.5-3.0 microM (200-1200 micrograms/L) CYN. To assess the usefulness of this assay, a range of toxic and nontoxic Cylindrospermopsis raciborskii extracts, including both laboratory strains and environmental samples, were assayed by protein synthesis inhibition. These CYN quantifications were then compared to quantifications obtained by high performance liquid chromatography (HPLC) and HPLC-tandem mass spectrometry (HPLCMS-MS). The results demonstrate that the protein synthesis inhibition assay correlates well with both HPLCMS-MS (r2 = 0.99) and HPLC (r2 = 0.97) quantifications. We conclude that this is an accurate and rapid assay for the measurement of cylindrospermopsin in cyanobacterial extracts. |
Keywords: | Reticulocytes Animals Rabbits Cyanobacteria Alkaloids Uracil Bacterial Toxins Protein Synthesis Inhibitors Biological Assay Chromatography, High Pressure Liquid Sensitivity and Specificity Dose-Response Relationship, Drug Mass Spectrometry Cyanobacteria Toxins |
DOI: | 10.1002/tox.1050 |
Published version: | http://dx.doi.org/10.1002/tox.1050 |
Appears in Collections: | Aurora harvest 2 Environment Institute publications Pharmacology publications |
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