title: Aldehyde oxidase 1 (AOX1) in human liver cytosols: quantitative characterization of AOX1 expression level and activity relationship.
abstract: Aldehyde oxidase 1 (AOX1) is a cytosolic enzyme highly expressed in liver and plays a key role in metabolizing drugs containing aromatic azaheterocyclic substituents. Rapid metabolism catalyzed by AOX1 can cause a drug to exhibit high clearance, low exposure, and hence decreased efficacy or even increased toxicity (if AOX1 generated metabolites are toxic). There is a need to develop the correlation between AOX1 expression levels and AOX1-substrate clearance. A fast, sensitive, and robust liquid chromatography-tandem mass spectrometry (LC-MS/MS) method was developed to quantify AOX1 in human liver cytosol for the first time. This LC-MS/MS method includes a straightforward ultrafiltration fractionation step and gives great selectivity and wide dynamic range (5.2 pM to 20.7 nM). The AOX1 levels in human liver cytosols of 20 donors were quantified using this method to investigate individual differences in AOX1 expression. No significant individual or gender differences in AOX1 levels were observed, although male donors exhibited a broader distribution than female donors (0.74-2.30 pmol/mg versus 0.74-1.69 pmol/mg, respectively). The AOX1 protein levels measured by LC-MS/MS were consistent with those measured by an enzyme-linked immunosorbent assay. Several donors have a normal AOX1 protein level but low enzyme activity, which might be due to cofactor deficiency, single nucleotide polymorphism, or homodimer dissociation. Cytosols from donors with chronic alcohol consumption had low AOX1-catalyzed carbazeran oxidation activities (<51 µl/min per milligram compared with a median of 455 µl/min per milligram), but preserved similar AOX1 protein expression levels (approximately 15% less than the median value).
authors.affiliation: Department of Pharmacokinetics, Dynamics, and Metabolism, Pfizer Worldwide Research and Development, Pfizer, Inc., Groton, Connecticut (C.F., L.D., X.H., C.S., M.D.T., R.S.O., H.Z.); and Scienion US, Inc., Monmouth Junction, New Jersey (M.S.).
authors.forename: Cexiong
authors.initials: C
authors.lastname: Fu
authors.forename: Li
authors.initials: L
authors.lastname: Di
authors.forename: Xiaogang
authors.initials: X
authors.lastname: Han
authors.forename: Cathy
authors.initials: C
authors.lastname: Soderstrom
authors.forename: Mark
authors.initials: M
authors.lastname: Snyder
authors.forename: Matthew D
authors.initials: MD
authors.lastname: Troutman
authors.forename: R Scott
authors.initials: RS
authors.lastname: Obach
authors.forename: Hui
authors.initials: H
authors.lastname: Zhang
mesh_terms: D000328:Adult; D000368:Aged; D042931:Aldehyde Oxidase / Q000737:chemistry* / Q000378:metabolism*; D002853:Chromatography, Liquid / Q000379:methods; D003600:Cytosol / Q000737:chemistry* / Q000201:enzymology / Q000378:metabolism*; D005260:Female; D006801:Humans; D008099:Liver / Q000737:chemistry / Q000201:enzymology / Q000378:metabolism*; D008297:Male; D008875:Middle Aged; D010449:Peptide Mapping / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods; D055815:Young Adult
chemical_list: C526436:AOX1 protein, human; D042931:Aldehyde Oxidase