Multiplicity of UDP-glucuronosyltransferases in fish. Purification and characterization of a phenol UDP-glucuronosyltransferase from the liver of a marine teleost, Pleuronectes platessa.
The aim of this work was to determine if a non-mammalian species had multiple UDP-glucuronosyltransferase (UDPGT) isoforms. At least six highly purified UDPGT isoenzymes were partially resolved by anion-exchange chromatography and UDP-hexanolamine-Sepharose 4B affinity chromatography from liver microsomes of a fish, the plaice. Q-Sepharose FF, chromatofocusing and affinity-chromatographic procedures were employed to separate and purify the phenol UDPGT isoform to apparent homogeneity. The purified enzyme conjugated 1-naphthol, but not bilirubin or steroids, and displayed a pI of 7.0 and a subunit molecular mass of 55 kDa. Bilirubin and testosterone UDPGT activities were more labile and, although purified over 200-fold, these preparations also contained the phenol UDPGT and had multiple polypeptides with molecular masses of 52-57 kDa. Antisera to rat bilirubin/phenol UDPGT and testosterone/phenol UDPGT isoforms cross-reacted strongly with the partially purified plaice UDPGT isoforms of molecular masses 52, 53 and 57 kDa and less strongly with phenol UDPGT 54 kDa and 56 kDa isoforms. Fish and mammalian UDPGTs therefore apparently possess a high degree of evolutionary conservation.
Clarke
D J
DJ
Department of Biochemical Medicine, University of Dundee, Ninewells Hospital and Medical School, U.K.
George
S G
SG
Burchell
B
B
D000818:Animals; D001663:Bilirubin / Q000378:metabolism; D015153:Blotting, Western; D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D005432:Flounder; D014453:Glucuronosyltransferase / Q000302:isolation & purification / Q000378:metabolism*; D007527:Isoenzymes / Q000302:isolation & purification / Q000378:metabolism*; D008862:Microsomes, Liver / Q000201:enzymology*; D009284:Naphthols / Q000378:metabolism; D010743:Phospholipids / Q000378:metabolism; D013256:Steroids / Q000378:metabolism; D013379:Substrate Specificity
D007527:Isoenzymes; D009284:Naphthols; D010743:Phospholipids; D013256:Steroids; C029350:1-naphthol; D014453:Glucuronosyltransferase; D001663:Bilirubin