Substrate characterisation of a recombinant sulfotransferase SULT1 and mRNA expression in chub (Leuciscus cephalus) tissues.
We have studied the role and regulation of sulfonation of xenobiotics and endogenous substrates in Leuciscus cephalus, an abundant and environmentally relevant freshwater fish. A sulfotransferase 1 (SULT1) cDNA and promoter region was cloned from chub liver and the cDNA expressed in Escherichia coli. The translated protein displayed 51% and 50% amino acid identity to mSULT1D1, hSULT1A1, respectively. We identified two conserved co-substrate binding motifs for 3'-phosphoadenosine 5'-phosphosulfate: RKGxxGDWKxxFT and YPKSGTxW. The recombinant SULT displayed a strong preference towards the isoflavones genistein (K(m)=1.7 microM) and daidzein (K(m)=4.4 microM) and lesser activity towards the endogenous substrate dopamine. Based on sequence identity and substrate preferences, the SULT was classified as SULT1,3. SULT1,3 mRNA expression was highest in the liver and kidney with low levels expressed in brain, gonad, and gill. Mature males displayed higher hepatic SULT1,3 mRNA expression compared to females. Analysis of the promoter region revealed several putative half palindromic estrogen response elements (ERE).
Assem
F L
FL
School of Biosciences, The University of Birmingham, Edgbaston, Birmingham, B15 2TT, UK.
Kirk
C J
CJ
Chipman
J K
JK
D000595:Amino Acid Sequence; D000818:Animals; D015239:Arylsulfotransferase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D001665:Binding Sites; D003001:Cloning, Molecular; D017124:Conserved Sequence; D003530:Cyprinidae / Q000235:genetics / Q000378:metabolism*; D018076:DNA, Complementary / Q000235:genetics; D015870:Gene Expression; D008099:Liver / Q000201:enzymology; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D013379:Substrate Specificity
D018076:DNA, Complementary; D012333:RNA, Messenger; D011994:Recombinant Proteins; D015239:Arylsulfotransferase