title: Hepatic and branchial glutathione S-transferases of two fish species: substrate specificity and biotransformation of microcystin-LR.
abstract: Liver and gills of roach (Rutilus rutilus) and silver carp (Hypophthalmichthys molitrix) were examined for glutathione S-transferases (GSTs) contents and their substrate specificity and capacity to biotransform microcystin-LR (MC-LR). GSTs and other glutathione (GSH) affine proteins were purified using a GSH-agarose matrix and separated by anionic chromatography (AEC). Substrate specificities were determined photometrical for 1-chloro-2,4-dinitrobenzene (CDNB), 1,2-dichloro-4-nitrobenzene (DCNB), 4-nitrobenzyl chloride (pNBC) and ethacrynic acid (ETHA). Biotransformation rate of MC-LR was determined by high performance liquid chromatography (HPLC). Roach exhibited different hepatic and branchial GST activities for used substrates (DNB, pNBC and DCNB) compared to silver carp but not for ethacrynic acid. It suggests that, both fish species have similar amount of pi and/or alpha class, which were the dominant GST classes in liver and gills. Gills of both fish species contained a higher number of GST isoenzymes, but with lower activities and ability of MC-LR biotransformation than livers. GST isoenzymes from roach had higher activity to biotransform MC-LR (conversion rate ranging up to 268 ng MC-LR min(-1) mL(-1) hepatic enzyme) than that isolated from silver carp. Without any prior contact to MC-LR or another GST inducer, roach seems to be better equipped for microcystin biotransformation than silver carp.
authors.affiliation: Faculty of Agriculture, University of South Bohemia in Ceské Budejovice, Ceské Budejovice, Czech Republic. setlik@zf.jcu.cz
authors.forename: I
authors.initials: I
authors.lastname: Setlíková
authors.forename: C
authors.initials: C
authors.lastname: Wiegand
mesh_terms: D000818:Animals; D001711:Biotransformation; D002347:Carps / Q000378:metabolism*; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D003530:Cyprinidae / Q000378:metabolism*; D004137:Dinitrochlorobenzene / Q000378:metabolism; D004976:Ethacrynic Acid / Q000378:metabolism; D005880:Gills / Q000201:enzymology*; D005982:Glutathione Transferase / Q000302:isolation & purification / Q000378:metabolism*; D007527:Isoenzymes / Q000302:isolation & purification / Q000378:metabolism; D008099:Liver / Q000201:enzymology*; D008387:Marine Toxins; D052998:Microcystins / Q000378:metabolism*; D009578:Nitrobenzenes / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D004137:Dinitrochlorobenzene; D007527:Isoenzymes; D008387:Marine Toxins; D052998:Microcystins; D009578:Nitrobenzenes; C027562:4-nitrobenzyl chloride; C028328:1,2-dichloro-4-nitrobenzene; D005982:Glutathione Transferase; C057862:cyanoginosin LR; D004976:Ethacrynic Acid