Proteomics approach on microcystin binding proteins in mouse liver for investigation of microcystin toxicity.
Microcystins (MC) produced by freshwater cyanobacteria are potent hepatotoxins. MC inhibit protein phosphatases (PP) 1 and 2A. MC and okadaic acid (OA), which is a similar PP inhibitor whereas it has a less affinity to PP1 than PP2A, behave similarly to primary culture hepatocytes, with inducements of phosphorylations of cytoskeleton, morphological changes and apoptosis. Although the distribution of OA in mouse liver was observed immunohistochemically, no OA injury was found. The purpose of this study was therefore to determine why only MC has specific toxicities on the liver. A systematic process of MC affinity chromatography and proteomics, using two-dimensional gel electrophoresis and MALDI-TOFMS, indicated the existence of some MC-binding proteins including the complexes of PP1, PP2A, and PP4 with their own regulatory subunits in mouse liver extracts. The competitive inhibition experiments using affinity chromatography with OA showed that two of the three protein complexes strongly interacted with OA, whereas only the complex of PP1 with the inhibitory subunit NIPP1 did not strongly interacted with OA. These results suggest that the PP1 complex is not related to the common behavior of MC and OA of primary culture hepatocytes, and is related to the specific hepatotoxicities of MC.
Imanishi
Susumu
S
Graduate School of Environmental and Human Sciences, Meijo University, Tempaku, Nagoya 468-8503, Japan. d6020004@ccmailg.meijo-u.ac.jp
Harada
Ken-ichi
K
D000818:Animals; D001427:Bacterial Toxins / Q000494:pharmacology*; D002352:Carrier Proteins / Q000037:antagonists & inhibitors / Q000187:drug effects; D022781:Hepatocytes / Q000187:drug effects*; D008099:Liver / Q000201:enzymology*; D008297:Male; D008387:Marine Toxins / Q000494:pharmacology*; D051379:Mice; D008807:Mice, Inbred BALB C; D052998:Microcystins; D019319:Okadaic Acid / Q000494:pharmacology; D010456:Peptides, Cyclic / Q000494:pharmacology*; D010749:Phosphoprotein Phosphatases / Q000037:antagonists & inhibitors / Q000187:drug effects*
D001427:Bacterial Toxins; D002352:Carrier Proteins; D008387:Marine Toxins; D052998:Microcystins; D010456:Peptides, Cyclic; D019319:Okadaic Acid; D010749:Phosphoprotein Phosphatases; C057862:cyanoginosin LR