Title: Role of Human Aldo-Keto Reductases in the Metabolic Activation of the Carcinogenic Air Pollutant 3-Nitrobenzanthrone.
Authors: Murray, Jessica R; Mesaros, Clementina A; Arlt, Volker M; Seidel, Albrecht; Blair, Ian A; Penning, Trevor M
Published In Chem Res Toxicol, (2018 11 19)
Abstract: 3-Nitrobenzanthrone (3-NBA) is a potent mutagen and suspected human carcinogen detected in diesel exhaust particulate and ambient air pollution. It requires metabolic activation via nitroreduction to promote DNA adduct formation and tumorigenesis. NAD(P)H:quinone oxidoreductase 1 (NQO1) has been previously implicated as the major nitroreductase responsible for 3-NBA activation, but it has recently been reported that human aldo-keto reductase 1C3 (AKR1C3) displays nitroreductase activity toward the chemotherapeutic agent PR-104A. We sought to determine whether AKR1C isoforms could display nitroreductase activity toward other nitrated compounds and bioactivate 3-NBA. Using discontinuous enzymatic assays monitored by UV-HPLC, we determined that AKR1C1-1C3 catalyze three successive two-electron nitroreductions toward 3-NBA to form the reduced product 3-aminobenzanthrone (3-ABA). Evidence of the nitroso- and hydroxylamino- intermediates were obtained by UPLC-HRMS. Km, kcat, and kcat/ Km values were determined for recombinant AKR1C and NQO1 and compared. We found that AKR1C1, AKR1C3, and NQO1 have very similar apparent catalytic efficiencies (8 vs 7 min-1 mM-1) despite the higher kcat of NQO1 (0.058 vs 0.012 min-1). AKR1C1-1C3 possess a Km much lower than that of NQO1, which suggests that they may be more important than NQO1 at the low concentrations of 3-NBA to which humans are exposed. Given that inhalation represents the primary source of 3-NBA exposure, we chose to evaluate the relative importance of AKR1C1-1C3 and NQO1 in human lung epithelial cell lines. Our data suggest that the combined activities of AKR1C1-1C3 and NQO1 contribute equally to the reduction of 3-NBA in A549 and HBEC3-KT cell lines and together represent approximately 50% of the intracellular nitroreductase activity toward 3-NBA. These findings have significant implications for the metabolism of nitrated polycyclic aromatic hydrocarbons and suggest that the hitherto unrecognized nitroreductase activity of AKR1C enzymes should be further investigated.
PubMed ID: 30406992
MeSH Terms: A549 Cells; Activation, Metabolic; Air Pollutants/analysis; Air Pollutants/metabolism*; Aldo-Keto Reductase Family 1 Member C3/antagonists & inhibitors; Aldo-Keto Reductase Family 1 Member C3/genetics; Aldo-Keto Reductase Family 1 Member C3/metabolism*; Benz(a)Anthracenes/analysis; Benz(a)Anthracenes/metabolism*; Biocatalysis; Cell Line; Chromatography, High Pressure Liquid; Humans; Mass Spectrometry; NAD(P)H Dehydrogenase (Quinone)/antagonists & inhibitors; NAD(P)H Dehydrogenase (Quinone)/genetics; NAD(P)H Dehydrogenase (Quinone)/metabolism; Protein Isoforms/antagonists & inhibitors; Protein Isoforms/genetics; Protein Isoforms/metabolism; Recombinant Proteins/genetics; Recombinant Proteins/metabolism