Quantification of diarrhetic shellfish toxins and identification of novel protein phosphatase inhibitors in marine phytoplankton and mussels.
Liquid chromatography (LC)-linked protein phosphatase 1/2A (PP-1/PP2A) bioassay was used to quantitatively identify diarrhetic shellfish toxins in marine phytoplankton (cultured and natural assemblages) and commercially available mussels. Using this approach, multiple protein phosphatase inhibitor profiles of varying composition were found in diarrhetic mussels from Holland and Canada. Based on LC elution positions and relative activity versus PP-1 and PP-2A, at least six inhibitors distinct from known diarrhetic shellfish toxins were identified and termed mussel phosphatase inhibitor (MPI) 19,22,23,25,33 and 42. The levels of these inhibitors, in okadaic acid equivalent units, varied from 100 pg to 3350 ng per g shellfish tissue. The combined levels of PP-1/2A inhibitors in all instances superseded that of okadaic acid/dinophysistoxin-1 and may contribute to the diarrhetic shellfish toxin profile of the contaminated mussels. The efficacy of LC-protein phosphatase bioassay was established for cultured phytoplankton where picogram levels of okadaic acid could be detected from microgram extracts of Prorocentrum lima. Analyses of plankton net tows from estuarine mussel culture sites in Eastern Canada revealed a heterogeneous population of protein phosphatase inhibitors, with dinophysistoxin-1 being most prevalent. This toxin was predominant for at least 2 months in mussel populations in the immediate vicinity of plankton sampling sites. The results are consistent with a hypothetical model in which marine bacteria, cyanobacteria and dinoflagellates combine to produce a variety of protein phosphatase inhibitors effective against signal transduction pathways in higher eukaryotes.
Luu
H A
HA
Department of Biochemistry, University of Alberta, Edmonton, Canada.
Chen
D Z
DZ
Magoon
J
J
Worms
J
J
Smith
J
J
Holmes
C F
CF
D000818:Animals; D049872:Bivalvia / Q000737:chemistry*; D002853:Chromatography, Liquid; D003967:Diarrhea / Q000139:chemically induced*; D004988:Ethers, Cyclic / Q000032:analysis; D008387:Marine Toxins / Q000032:analysis*; D019319:Okadaic Acid / Q000031:analogs & derivatives; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors*; D010839:Phytoplankton / Q000737:chemistry*; D054645:Protein Phosphatase 1; D011714:Pyrans / Q000032:analysis; D011817:Rabbits
D004988:Ethers, Cyclic; D008387:Marine Toxins; D011714:Pyrans; D019319:Okadaic Acid; C051904:dinophysistoxin 1; D010749:Phosphoprotein Phosphatases; D054645:Protein Phosphatase 1