Stereoselective sulfoxidation by human flavin-containing monooxygenase. Evidence for catalytic diversity between hepatic, renal, and fetal forms.
The stereoselective formation of p-tolyl methyl sulfoxide from the corresponding sulfide has been examined in detergent-solubilized human adult liver, adult kidney, and fetal liver microsomes, in order to compare the functional activities of human flavin-containing monooxygenase(s). Solubilization with detergent was performed to eradicate the contribution that cytochrome P-450 would make to the net stereochemistry. Consistent with studies in experimental animal livers, solubilized human fetal liver and adult kidney microsomes formed (R)-p-tolyl methyl sulfoxide in greater than 86% enantiomeric excess. These enzyme activities were sensitive to methimazole inhibition and were markedly thermolabile in the absence of NADPH, attributes that are consistent with a flavin-containing monooxygenase-mediated process. However, solubilized adult human liver microsomes displayed little stereoselectivity (0-40% enantiomeric excess) for the formation of (R)-p-tolyl methyl sulfoxide, although this reaction also displayed several of the characteristics of a flavin-containing monooxygenase-dependent process, including sensitivity to methimazole inhibition and NADPH protection against heat inactivation. Furthermore, this lack of stereoselectivity was not attenuated by the inclusion of activated oxygen scavengers in reaction mixtures. Human tissue metabolite profiling was further studied by using the ethyl, propyl, and isopropyl p-tolyl sulfides. Parallel changes in product stereochemistry as a function of increasing steric bulk were observed with the fetal liver and adult kidney tissue, whereas an anomalous profile was again observed with adult human liver. These data are consistent with the presence of functionally discrete complements of the flavin-containing monooxygenase in detergent-solubilized adult and fetal human liver microsomes.
Sadeque
A J
AJ
Department of Medicinal Chemistry, University of Washington, Seattle 98195.
Eddy
A C
AC
Meier
G P
GP
Rettie
A E
AE
D000328:Adult; D002374:Catalase / Q000494:pharmacology; D003902:Detergents / Q000494:pharmacology; D005333:Fetus / Q000201:enzymology*; D006801:Humans; D066298:In Vitro Techniques; D007668:Kidney / Q000201:enzymology*; D008861:Microsomes / Q000201:enzymology; D008862:Microsomes, Liver / Q000201:enzymology*; D010105:Oxygenases / Q000494:pharmacology*; D013237:Stereoisomerism; D013454:Sulfoxides / Q000378:metabolism*; D013482:Superoxide Dismutase / Q000494:pharmacology
D003902:Detergents; D013454:Sulfoxides; D002374:Catalase; D010105:Oxygenases; C020066:dimethylaniline monooxygenase (N-oxide forming); D013482:Superoxide Dismutase