Abstract:
:Uridine diphosphate glucuronosyltransferases (UGTs) are important phase II detoxification enzymes. Despite the expression of UGT proteins in many species, previous results have suggested that simians represent the most appropriate animal model to study the glucuronidation of steroids in extrahepatic steroid target tissues. Northern blot analysis using a pool of human UGT2B cDNA probes demonstrated the expression of homologous UGT2B transcripts in several tissues including the liver, kidney, adrenal, breast, testis, and prostate of the cynomolgus monkey (Macacafascicularis). Western blot analyses using a polyclonal antibody raised against human UGT2B17 protein also demonstrated expression of homologous UGT2B proteins in monkey tissues. cDNA libraries were constructed from monkey liver and prostate mRNA and a novel UGT2B cDNA, UGT2B9, was isolated from both libraries. The UGT2B cDNA from the prostate library is 2,648 bp in length and contains an open reading frame of 1,587 bp encoding a protein of 529 residues. In vitro transcription/translation of the cDNA clone produced a protein of 52 kD. The UGT2B9 cDNA clone was transfected into HK293 cells and a stable cell line expressing UGT2B9 protein was established. The activity of UGT2B9 was tested with over 60 compounds and was demonstrated to be active on C18, C19, and C21 steroids, bile acids, and several xenobiotics including eugenol, 1-naphthol, and p-nitrophenol. Kinetic analysis revealed that UGT2B9 glucuronidates steroids with high affinity and efficiency with Km values of 0.2, 3.2, 0.2, and 1.8 microM for dihydrotestosterone, testosterone, androsterone, and 1,3,5,10-estratrien-3,4-diol-17-one, respectively. It is apparent that this simian UGT2B enzyme is specific for more different classes of steroids than any other UGT enzyme characterized to date, and may be related to the high plasma levels of glucuronidated C19 steroids found in the cynomolgus monkey.
journal_name
DNA Cell Bioljournal_title
DNA and cell biologyauthors
Bélanger G,Beaulieu M,Lévesque E,Hum DW,Bélanger Adoi
10.1089/dna.1997.16.1195subject
Has Abstractpub_date
1997-10-01 00:00:00pages
1195-205issue
10eissn
1044-5498issn
1557-7430journal_volume
16pub_type
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