Document Detail


Identification and characterization of hydrophobic microcystins in Canadian freshwater cyanobacteria.
MedLine Citation:
PMID:  8146867     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Hepatotoxic microcystins produced by cyanobacteria in freshwater lakes represent a significant health hazard to humans and agricultural livestock. Liquid chromatography (LC)-linked protein phosphatase (PPase) bioassay analysis of blooms of Microcystis aeruginosa produced in a Canadian drinking water lake identified several PPase inhibitors with significantly greater hydrophobicity than microcystin-LR, based on their retention time on C18 reverse phase LC columns. Seven PPase inhibitors were purified to homogeneity by bioassay-guided fractionation involving Sephadex LH-20 chromatography and two-step reverse phase at pH 6.5 and 2.0. One of the PPase inhibitors, isolated in a final yield of 1.5 micrograms/g lyophilized cyanobacteria, was identified as microcystin-LL by amino acid analysis and mass spectrometry. A further PPase inhibitor (20 ng/g cyanobacteria) was identified as microcystin-LL but with D-Ala replaced by an unknown amino acid. Four PPase inhibitors (< 20 ng/g cyanobacteria) were characterized by amino acid analysis and identified as microcystin-LV, -LM, -LF and -LZ (where Z represents an unknown hydrophobic amino acid). A further microcystin was also identified (< 10 ng/g cyanobacteria) in which arginine was apparently absent. The biological activity of the seven microcystins as inhibitors of the catalytic subunit of protein phosphatase-1 (PP-1c) was compared with microcystin-LR and motuporin (a hydrophobic analogue of nodularin). All of the compounds inhibited PP-1c with IC50 values of 0.06-0.4 nM, consistent with their identification as microcystins. These findings further demonstrate the applicability of a sensitive PPase bioassay for the identification of variant microcystins in the natural environment.
Authors:
M Craig; T L McCready; H A Luu; M A Smillie; P Dubord; C F Holmes
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Toxicon : official journal of the International Society on Toxinology     Volume:  31     ISSN:  0041-0101     ISO Abbreviation:  Toxicon     Publication Date:  1993 Dec 
Date Detail:
Created Date:  1994-05-05     Completed Date:  1994-05-05     Revised Date:  2007-11-15    
Medline Journal Info:
Nlm Unique ID:  1307333     Medline TA:  Toxicon     Country:  ENGLAND    
Other Details:
Languages:  eng     Pagination:  1541-9     Citation Subset:  IM    
Affiliation:
Department of Biochemistry, University of Alberta, Edmonton, Canada.
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MeSH Terms
Descriptor/Qualifier:
Amino Acids / analysis
Canada
Chromatography, Liquid
Cyanobacteria / chemistry*
Eutrophication / physiology
Microcystins
Peptides, Cyclic / analysis*,  chemistry,  pharmacology
Phosphoprotein Phosphatases / analysis,  antagonists & inhibitors
Protein Phosphatase 1
Spectrophotometry, Ultraviolet
Water Microbiology
Water Supply / analysis
Chemical
Reg. No./Substance:
0/Amino Acids; 0/Microcystins; 0/Peptides, Cyclic; 77238-39-2/microcystin; EC 3.1.3.16/Phosphoprotein Phosphatases; EC 3.1.3.16/Protein Phosphatase 1

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