Role of Fish Hepatic Aldehyde Oxidase in Oxidative in vitro Metabolism of Phenanthridine Heterocyclic Aromatic Compound

Authors: Khaled S. Al Salhen

Abstract:

Aldehyde oxidase is molybdo-flavoenzyme involved in the oxidation of hundreds of endogenous and exogenous and N-heterocyclic compounds and environmental pollutants. Uncharged N-heterocyclic aromatic compounds such phenanthridine are commonly distributed pollutants in soil, air, sediments, surface water and groundwater, and in animal and plant tissues. Phenanthridine as uncharged N-heterocyclic aromatic compound was incubated with partially purified aldehyde oxidase from rainbow trout fish liver. Reversed-phase HLPC method was used to separate the oxidation products from phenanthridine and the metabolite was identified. The 6(5H)-phenanthridinone was identified the major metabolite by partially purified aldehyde oxidase from fish liver. Kinetic constant for the oxidation reactions were determined spectrophotometrically and showed that this substrate has a good affinity (K_m = 78 ± 7.6µM) for hepatic aldehyde oxidase, will be a significant pathway. This study confirms that partially purified aldehyde oxidase from fish liver is indeed the enzyme responsible for the in vitro production 6(5H)-phenanthridinone metabolite as it is a major metabolite by mammalian aldehyde oxidase, coupled with a relatively high oxidation rate (0.77± 0.03 nmol/min/mg protein). In addition, the kinetic parameters of hepatic fish aldehyde oxidase towards the phenanthridine substrate indicate that in vitro biotransformation by hepatic fish aldehyde oxidase will be a significant pathway. This study confirms that partially purified aldehyde oxidase from fish liver is indeed the enzyme responsible for the in vitro production 6(5H)-phenanthridinone metabolite as it is a major metabolite by mammalian aldehyde oxidase.

Keywords: Aldehyde oxidase, Fish, Phenanthridine, Specificity.

Digital Object Identifier (DOI): doi.org/10.5281/zenodo.1091236

References:

[1] C. Beedham, "Enzyme Systems that Metabolise Drugs and Other Xenobiotics”. CostasIoannides (Ed). Copyright © 2001John Wiley & Sons Ltd.
[2] S. Kitamura, K. Sugihara, and S. Ohta, "Drug-Metabolizing Ability of Molybdenum Hydroxylases,” Drug Metab.Pharmacokinet. Vol. 21, no. 2, pp. 83-98, 2006.
[3] C. Beedham, "Xanthine Oxidoreductaseand Aldehyde Oxidase”. (Second ed. Chapter 4.10): Comprehensive Toxicology. Copyright © 2010 Elsevier Ltd. All rights reserved.
[4] E. Garattini, and M. Terao, "The Role of Aldehyde Oxidase in Drug Metabolism,” Expert.Opin. Drug Metab. Toxicol.Vol. 8, no. 4, pp. 487-503, 2012.
[5] E. Garattini, M. Fratelli, and M. Terao, "The Mammalian Aldehyde Oxidase Gene Family,” Hum. Genomics. Vol. 4, no. 2, pp. 119-130, 2009.
[6] D. Schlenk, "Occurrence of Flavin-Containing Monooxygenasesin Non-Mammalian Eukaryotic Organisms,” Comparative Biochemistry and Physiology Part C: Pharmacology Toxicology and Endocrinology, Vol. 121, no. 1-3, pp. 185-195 1998.
[7] M. Kurosaki, M. Bolis, M. Fratelli, M. Barzago, L. Pattini, G. Perretta, M. Terao, and E. Garattini, "Structure and Evolution of Vertebrate Aldehyde Oxidases: From Gene Duplication to Gene Suppression,” Cellular and Molecular Life Sciences. Vol. 70, no. 10, pp.1807-1830, 2013.
[8] K. Tatsumi, S. Kitamura, M. Kato, and K. Hiraoka, "Metabolism of Sodium Nifurstyrenate, a Veterinary Antimicrobial Nitrofuranin Animals and Fish,” Drug Metab.Dispos. Vol, 20, no. 2, pp. 226-233, 1992.
[9] S. Kitamura, and K. Tatsumi, "1-Nitropyrene-Metabolizing Activities of Fish Liver Preparations,” Bull. Environ. Contam. Toxicol. Vol 58, no. 3, pp. 448–455 1997.
[10] O. Ueda, S. Kitamura, and S. Ohta, "Metabolism of 2-Nitrofluorene, an Environmental Pollutant, by Liver Preparations of Sea Bream, PagrusMajor,” Xenobiotica.Vol. 32, no. 8, pp. 667-682, 2002.
[11] M. R. Lakshmanan, C. S. Vaidyanathan, and H. R. Cama, "Oxidation of Vitamin A1 Aldehyde and Vitamin A2 Aldehyde to the Corresponding Acids by Aldehyde Oxidase from Different Species,” Biochem J. Vol. 90, no. 3, pp. 569–573, 1964.
[12] W. Brack, and K. Schirmer, "Effect-Directed Identification of Oxygen and Sulfur Heterocyclesas Major Polycyclic Aromatic Cytochrome P4501A-Inducers in Contaminated Sediment,” Environmental Science &Technology.Vol. 37, no. 14, pp. 3062-3070, 2003.
[13] S. Meyer, and H. Steinhart, "Effects of Heterocyclic PAHs (N, S and O) on the Biodegradation of Typical Tar Oil PAHs in a Soil/Compost Mixture,” Chemosphere.Vol. 40, pp. 359-367, 2000.
[14] S. Peddinghaus, M. Brinkmann, K. Bluhm, A. Sagner, G. Hinger, T. Braunbeck, A. Eisentraeger, A. Tiehm, H. Hollert, and S. H. Keiter, "Quantitative Assessment of the Embryotoxic Potential of NSO-Heterocyclic Compounds Using Zebra Fish (Daniorerio),” Reproductive Toxicology. Vol. 33, no. 2, pp. 224-232, 2012.
[15] M. G. Barron, R. Heintz, and S. D. Rice, "Relative Potency of PAHs and Heterocyclesas Aryl Hydrocarbon Receptor Agonists in Fish,” Mar. Environ. Res. Vol. 58, no. 2-5, pp. 95-100, 2004.
[16] D. K. Jung, T. Klaus, and K. Fent, "Cytochrome P450 Induction by Nitrated Polycyclic Aromatic Hydrocarbons, Azaarenes, and Binary Mixtures in Fish Hepatoma Cell Line PLHC-1,” Environmental Toxicology and Chemistry. Vol. 20, no. 1, pp. 149-159, 2001.
[17] M. Feldmannova, K. Hilscherova, B. Marsalek, and L. Blaha, "Effects of N-Heterocyclic Polyaromatic Hydrocarbons on Survival, Reproduction, and Biochemical Parameters in Daphnia magna,” Environmental Toxicology.Vol. 21 no. 4, pp. 425-431 2006.
[18] I. Sovadinova, L. Blaha, J. Janosek, K. Hilscherova, J. P. Giesy, P. D. Jones, and I. Holoubek, Cytotoxicity and Aryl Hydrocarbon Receptor-Mediated Activity of N-Heterocyclic Polycyclic Aromatic Hydrocarbons: Structure-Activity Relationships, Environmental Toxicology and Chemistry. Vol. 25, no. 5, pp. 1291-1297, 2006.
[19] P. L. Van Vlaardingen, W. J. Steinhoff, P. De Voogt, and W. A. Admiraal, "Property Toxicity Relationships of Azaarenesto the Green Alga Scenedesmusacuminatus,” Enviro. Tpxicol. Chem. Vol. 15, no. 11, pp. 2035-2045, 1996.
[20] W. J. Catallo, "Transformation of N-, O-, and S-Heterocycles(NOSHs) in Estuarine Sediments: Effects of Redox Potential and Sediment Particle Size,” Chemosphere. Vol. 133, pp. 2543-2563, 1996.
[21] M. Machala, J. Vondrácˇek, L. Bláha, M. Cigánek, and J. Neca J, "Aryl Hydrocarbon Receptor-Mediated Activity of Mutagenic PAHs Determined Usingin vitroReporter Gene Assay,” Mutat Res. Genet. Toxicol. Environ. Mutat. Vol. 497, no. 1-2, pp. 49-62, 2001.
[22] H. Y. Chen, and M. R. Preston, "Gas/Particle Partitioning Behaviourof Azaarenesin an Urban Atmosphere,” Environmental Pollution.Vol. 97, no. 1-2, pp. 169-174, 1997.
[23] E. A. J. Bleeker, L. Noor, P. Kraak, P. deVoogt, and W. Admiraal, "Comparative Metabolism of Phenanthridine by Carp (Cyprinuscarpio) and Midge Larvae (Chironomusriparius),” Environmental Pollution. Vol. 112, no. 1, pp. 11-17, 2001.
[24] E. J. LaVoie, A. Shigematsu, and D. Hoffmann, "Metabolites of Phenanthridine Formed by Rat Liver Homogenate,”DrugMetab. Dispos. Vol. 13, no. 1, pp. 71-75, 1985.
[25] C. Johnson, C. Beedham, J. G. Stell J, "Elevation of Molybdenum Hydroxylase Levels in Rabbit Liver after Ingestion of Phthalazineor Its Hydroxylated Metabolite,” Biochem. Pharmacol. Vol. 33, no. 22, pp. 3699-3705, 1984.
[26] P. K. Smith, R. I. Krohn, G. T. Hermanson, A. K. Mallia, F. H. Gartner, M. D. Provenzano, "Measurement of Protein Using Bicinchoninic Acid, "Anal. Biochem. Vol. 150, no. 1, pp. 76-85, 1985.
[27] B. Houston, K. E. Kenworthy, and A. Galetin, "Typical and Atypical Enzyme Kinetics, in Drug Metabolism Enzymes: Cytochrome P450 and Other Enzymes in Drug Discovery and Development,” Netherlands and New York: Fontis Media .A. and Marcel Dekker Inc, 2003.
[28] C. Beedham, "Molybdenum Hydroxylases: Biological Distribution and Substrate-Inhibitor Specificity,” Prog Med. Chem. Vol. 24, pp. 127-185, 1987.
[29] M. Terao, M. Kurosaki, G. Saltini, S. Demontis, M. Marini, M. Salmona, "Cloning of the cDNAsCoding for Two Novel Molybdo-Flavoproteins Showing High Similarity with Aldehyde Oxidase and Xanthine Oxidoreductase,” J. Biol. Chem. Vol. 275, pp. 30690-30700, 2000.
[30] S. Pirouzpanah, M. R. Rashidi, A. Delazar, S. V. Razavieh, and A. Hamidi, "Inhibitory Effects of Rutagraveolens L. Extract on Guinea Pig Liver Aldehyde Oxidase,” Chem. Pharm. Bull. (Tokyo). Vol. 54, no. 1,pp. 9-13, 2006.
[31] M. H. Sorouraddin, E. Fooladi, A. Naseri, M. R. Rashidi, "A Novel Spectrophotometric Method for Determination of Kinetic Constants of Aldehyde Oxidase Using Multivariate Calibration Method,” J. Biochem. Biophys. Methods. Vol. 70, no. 6, pp. 999-1005, 2009.
[32] C. Stubley, and J. G. Stell, "Investigation of the Substrate-Binding Site of Aldehyde Oxidase,” Journal of Pharmacy and Pharmacology.Vol. 32, no.S1, pp. 51, 1980.
[33] M. R. Rashidi, A. J. Smith, S. E. Clarke, C. Beedham, "In vitroOxidation of Famciclovirand 6-Deoxypenciclovir by Aldehyde Oxidase from Human, Guinea Pig, Rabbit, and Rat Liver,” Drug Metab. Dispos.Vol. 25, no. 7, pp. 805-813, 1997.
[34] E. Garattini, M. Fratelli, and M. Terao, "The Mammalian Aldehyde Oxidase Gene Family,” Hum. Genomics. Vol. 4, no. 2, pp. 119-130, 2009.
[35] M. Kurosaki, M. Bolis, M. Fratelli, M. M. Barzago, L. Pattini, G. Perretta, M. Terao, and E. Garattini, "Structure and Evolution of Vertebrate Aldehyde Oxidases: From Gene Duplication to Gene Suppression,” Cellular and Molecular Life Sciences. Vol. 70, no.10, pp. 1807-1830, 2013.
[36] C. Beedham, "Molybdenum Hydroxylases as Drug Metabolising Enzymes,” Drug Metab. Rev. Vol. 16, no. 1-2, pp. 111-156, 1985.
[37] G. I. Panoutsopoulos, D. Kouretas, E. G. Gounaris, and C. Beedham, "Enzymatic oxIdation of Vanillin, Isovanillinand Protocatechuic Aldehyde with Freshly Prepared Guinea Pig Liver Slices,” Cell Physiol. Biochem. Vol. 15, no. 1-4, pp. 89-98, 2005.
[38] G.I. Panoutsopoulos, D. Kouretas, E. G. Gounaris, and C. Beedham, "Kinetics and Specificity of Guinea Pig Liver Aldehyde Oxidase and Bovine Milk Xanthine Oxidase towards Substituted Benzaldehydes” Acta. Biochim. Pol. Vol. 51, no. 13, pp. 649-463, 2004.