Document Detail


Cell-free protein synthesis inhibition assay for the cyanobacterial toxin cylindrospermopsin.
MedLine Citation:
PMID:  11594027     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
S M Froscio; A R Humpage; P C Burcham; I R Falconer
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Environmental toxicology     Volume:  16     ISSN:  1520-4081     ISO Abbreviation:  Environ. Toxicol.     Publication Date:  2001 Oct 
Date Detail:
Created Date:  2001-10-11     Completed Date:  2002-02-05     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  100885357     Medline TA:  Environ Toxicol     Country:  United States    
Other Details:
Languages:  eng     Pagination:  408-12     Citation Subset:  IM    
Affiliation:
Department of Clinical and Experimental Pharmacology, University of Adelaide, Adelaide, South Australia 5005, Australia.
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MeSH Terms
Descriptor/Qualifier:
Animals
Biological Assay / methods
Chromatography, High Pressure Liquid
Cyanobacteria / chemistry*
Dose-Response Relationship, Drug
Mass Spectrometry
Protein Synthesis Inhibitors / analysis*,  pharmacology*
Rabbits
Reticulocytes / enzymology
Sensitivity and Specificity
Uracil / analogs & derivatives*,  analysis*,  pharmacology*
Chemical
Reg. No./Substance:
0/Protein Synthesis Inhibitors; 143545-90-8/cylindrospermopsin; 66-22-8/Uracil

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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