Highly sensitive detection and discrimination of LR and YR microcystins based on protein phosphatases and an artificial neural network.
The inhibition characteristics of three different protein phosphatases by three microcystin (MC) variants--LR, YR, and RR--were studied. The corresponding K (I) for each enzyme-MC couple was calculated. The toxicity of MC varies in the following order: MC-LR > MC-YR > MC-RR. The sensitivity of the enzymes increased in the following order: mutant PP2A < mutant PP1 < natural PP2A. The best limit of detection obtained was 21.2 pM MC-LR using the most sensible enzyme. Methanol, ethanol, and acetonitrile up to 2 % (v/v) may be used in inhibition measurements. An artificial neural network (ANN) was used to discriminate two MC variants--LR and YR--using the differences in inhibition percentages measured with mutant PP1 and natural PP2A. The ANN is able to analyze mixtures with concentrations ranging from 8 to 98 pM MC-LR and 31 to 373 pM MC-YR.
Laboratoire IMAGES EA 4218, University of Perpignan Via Domitia, Perpignan, France.
O I
OI
Covaci
A
A
Sassolas
G A
GA
Alonso
R
R
Muñoz
G L
GL
Radu
B
B
Bucur
J-L
JL
Marty
D000818:Animals; D003001:Cloning, Molecular; D004791:Enzyme Inhibitors / Q000032:analysis* / Q000737:chemistry; D004926:Escherichia coli; D006801:Humans; D007527:Isoenzymes / Q000737:chemistry / Q000235:genetics; D007700:Kinetics; D057230:Limit of Detection; D008387:Marine Toxins; D052998:Microcystins / Q000032:analysis* / Q000737:chemistry; D009154:Mutation; D016571:Neural Networks, Computer; D010456:Peptides, Cyclic / Q000032:analysis* / Q000737:chemistry; D054645:Protein Phosphatase 1 / Q000737:chemistry* / Q000235:genetics; D054648:Protein Phosphatase 2 / Q000737:chemistry* / Q000235:genetics; D011817:Rabbits; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D012997:Solvents
D004791:Enzyme Inhibitors; D007527:Isoenzymes; D008387:Marine Toxins; D052998:Microcystins; D010456:Peptides, Cyclic; D011994:Recombinant Proteins; D012997:Solvents; C065685:microcystin YR; C063855:microcystin RR; D054645:Protein Phosphatase 1; D054648:Protein Phosphatase 2; C057862:cyanoginosin LR