Purification of macaque liver flavin-containing monooxygenase: a form of the enzyme related immunochemically to an isozyme expressed selectively in adult human liver.
A microsomal flavin-containing monooxygenase (FMO) was purified 77-fold from macacque liver microsomes on the basis of its methyl p-tolyl sulfoxidase activity. Sequential chromatography on anion- and cation-exchangers, lauryl-Sepharose and 2',5'-ADP-Sepharose provided a purified preparation which exhibited an apparent molecular mass of 59 kDa and a pI of 8.3. N-terminal amino-acid sequencing revealed the characteristic Gly-X-Gly-X-X-Gly consensus sequence for the putative FAD-binding domain of microsomal FMO. In marked contrast to the well-characterized hepatic and pulmonary forms present in experimental animals, the macacque liver enzyme displayed stereoselectivity for sulfoxidation of p-tolyl methyl sulfide on the pro-S rather than the pro-R face of the substrate. Polyclonal antibodies raised against the macacque liver form exhibited little or no cross-reactivity with major purified forms of the enzyme isolated from rabbit liver, guinea-pig liver or rabbit lung. Anti-macacque liver FMO did not cross-react with human fetal liver or adult kidney microsomes, but did recognize a 59 kDa constituent of human adult liver microsomes. The intensity of this immunoreactive 59 kDa band correlated well with human liver microsomal N,N-dimethylaniline N-oxygenase activity. We conclude that human adult liver selectively expresses a microsomal FMO which is functionally and immunochemically distinct from the FMO form(s) present in human fetal liver and adult kidney, and from the major hepatic and pulmonary forms present in common laboratory animals.
Sadeque
A J
AJ
Department of Medicinal Chemistry, University of Washington, Seattle 98195.
Thummel
K E
KE
Rettie
A E
AE
D000328:Adult; D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology; D003429:Cross Reactions; D006168:Guinea Pigs; D006801:Humans; D007668:Kidney / Q000201:enzymology; D008168:Lung / Q000201:enzymology; D008251:Macaca; D008862:Microsomes, Liver / Q000201:enzymology*; D008969:Molecular Sequence Data; D010105:Oxygenases / Q000032:analysis / Q000276:immunology / Q000302:isolation & purification*; D011817:Rabbits
D000906:Antibodies; D010105:Oxygenases; C020066:dimethylaniline monooxygenase (N-oxide forming)