cDNA expression studies of rat liver aryl sulphotransferase.
A cDNA encoding an isoenzyme of rat liver aryl sulphotransferase was isolated from a rat liver bacteriophage Lambda gt 11 library by the polymerase chain reaction technique. The resulting cDNA was functionally expressed in COS-7 cells and characterised by determining the sulphating capacity of the cells with a range of substrates. The COS-expressed enzyme catalysed the sulphation of both phenol and dopamine with Kms of the same order as those obtained for the high affinity isozyme in rat liver cytosol, while low activity was observed with tyrosine methyl ester. The common food additive vanillin was also a good substrate for sulphate conjugation. The sulphation of vanillin catalysed by the COS-expressed enzyme was consistent with a single enzyme system, in contrast, the kinetics of the reaction catalysed by cytosolic sulphotransferase indicated that vanillin was sulphated by more than one isozyme.
Cruickshank
D
D
School of Pharmacy, University of South Australia, Adelaide.
Sansom
L N
LN
Veronese
M E
ME
Mojarrabi
B
B
McManus
M E
ME
Zhu
X
X
D000818:Animals; D015239:Arylsulfotransferase / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D001547:Benzaldehydes / Q000378:metabolism; D002460:Cell Line; D003600:Cytosol / Q000201:enzymology; D004247:DNA / Q000235:genetics*; D004298:Dopamine / Q000378:metabolism; D015723:Gene Library; D007527:Isoenzymes / Q000235:genetics*; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D008297:Male; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D019800:Phenol; D010636:Phenols / Q000378:metabolism; D016133:Polymerase Chain Reaction / Q000379:methods; D051381:Rats; D017208:Rats, Wistar; D013379:Substrate Specificity; D014162:Transfection
D001547:Benzaldehydes; D007527:Isoenzymes; D009838:Oligodeoxyribonucleotides; D010636:Phenols; D019800:Phenol; D004247:DNA; C100058:vanillin; D015239:Arylsulfotransferase; D004298:Dopamine