Radiation inactivation analysis of microsomal UDP-glucuronosyltransferases catalysing mono- and diglucuronide formation of 3,6-dihydroxybenzo(a)pyrene and 3,6-dihydroxychrysene.
Indirect evidence has suggested that multiple subunits of microsomal UDP-glucuronosyltransferases (UGTs) are involved in diglucuronide formation of diphenols of polycyclic aromatic hydrocarbons (Bock et al., Mol Pharmacol 42: 613-618, 1992). To substantiate this suggestion functional target sizes of UGTs catalysing these reactions were determined in microsomes in situ by radiation inactivation analysis. Target sizes of UGTs catalysing the glucuronidation of 1-naphthol and 6-hydroxychrysene were found to be 91 +/- 29 and 120 +/- 27 kDa, respectively. However, target sizes for mono- and diglucuronide formation of 3,6-dihydroxybenzo(a)pyrene were 118 +/- 33 and 218 +/- 24 kDa, respectively. Similarly, using 3,6-dihydroxychrysene as substrate target sizes of 109 +/- 21 and 101 +/- 23 kDa were found for 6-O-monoglucuronide and 3-O-monoglucuronide formation and a target size of 192 +/- 34 kDa observed for diglucuronide formation. Based on subunit molecular masses of 50-60 kDa for UGTs, these results suggest that UGTs involved in monoglucuronide formation of phenols may function as dimers. In contrast, UGTs involved in diglucuronide formation of diphenols of polycyclic aromatic hydrocarbons may function as tetramers in microsomes in situ.
Institute of Toxicology, University of Tübingen, Germany.
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Gschaidmeier
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Bock
D000201:Activation Analysis; D000818:Animals; D001580:Benzopyrenes / Q000378:metabolism*; D002911:Chrysenes / Q000378:metabolism*; D005965:Glucuronates / Q000378:metabolism*; D014453:Glucuronosyltransferase / Q000737:chemistry / Q000378:metabolism*; D006873:Hydroquinones / Q000378:metabolism*; D008297:Male; D008748:Methylcholanthrene; D008862:Microsomes, Liver / Q000201:enzymology*; D009284:Naphthols / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar
C090230:3,6-dihydroxychrysene; D001580:Benzopyrenes; D002911:Chrysenes; D005965:Glucuronates; D006873:Hydroquinones; D009284:Naphthols; C029350:1-naphthol; C087203:6-hydroxychrysene; D008748:Methylcholanthrene; C054191:benzo(a)pyrene-3,6-quinol; D014453:Glucuronosyltransferase