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Laurieri, Nicola; Dairou, Julien; Egleton, James E.; Stanley, Lesley A.; Russell, Angela J.; Dupret, Jean-Marie; Sim, Edith; Rodrigues-Lima, Fernando (2014)
Publisher: Public Library of Science
Journal: PLoS ONE
Languages: English
Types: Article
Subjects: Enzyme Chemistry, Biotechnology, Research Article, Biology and Life Sciences, Developmental Biology, Enzymes, Medicine, Basic Cancer Research, Cancer Risk Factors, Birth Defects, Oncology, Enzymology, alliedhealth, Cofactors (Biochemistry), Q, R, Morphogenesis, Science, Biochemistry, Medicine and Health Sciences, Embryology, Small Molecules
Acetyl Coenzyme A-dependent N-, O- and N,O-acetylation of aromatic amines and hydrazines by arylamine N-acetyltransferases is well characterised. Here, we describe experiments demonstrating that human arylamine N-acetyltransferase Type 1 and its murine homologue (Type 2) can also catalyse the direct hydrolysis of acetyl Coenzyme A in the presence of folate. This folate-dependent activity is exclusive to these two isoforms; no acetyl Coenzyme A hydrolysis was found when murine arylamine N-acetyltransferase Type 1 or recombinant bacterial arylamine N-acetyltransferases were incubated with folate. Proton nuclear magnetic resonance spectroscopy allowed chemical modifications occurring during the catalytic reaction to be analysed in real time, revealing that the disappearance of acetyl CH 3 from acetyl Coenzyme A occurred concomitantly with the appearance of a CH 3 peak corresponding to that of free acetate and suggesting that folate is not acetylated during the reaction. We propose that folate is a cofactor for this reaction and suggest it as an endogenous function of this widespread enzyme. Furthermore, in silico docking of folate within the active site of human arylamine N-acetyltransferase Type 1 suggests that folate may bind at the enzyme’s active site, and facilitate acetyl Coenzyme A hydrolysis. The evidence presented in this paper adds to our growing understanding of the endogenous roles of human arylamine N-acetyltransferase Type 1 and its mouse homologue and expands the catalytic repertoire of these enzymes, demonstrating that they are by no means just xenobiotic metabolising enzymes but probably also play an important role in cellular metabolism. These data, together with the characterisation of a naphthoquinone inhibitor of folate-dependent acetyl Coenzyme A hydrolysis by human arylamine N-acetyltransferase Type 1/murine arylamine N-acetyltransferase Type 2, open up a range of future avenues of exploration, both for elucidating the developmental role of these enzymes and for improving chemotherapeutic approaches to pathological conditions including estrogen receptor-positive breast cancer.
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    • 1. Butcher NJ, Minchin RF (2012) Arylamine N-acetyltransferase 1: a novel drug target in cancer development. Pharmacol Rev 64: 147-165.
    • 2. Sim E, Fakis G, Laurieri N, Boukouvala S (2012) Arylamine N-acetyltransferases-from drug metabolism and pharmacogenetics to identification of novel targets for pharmacological intervention. Adv Pharmacol 63: 169-205.
    • 3. Sim E, Walters K, Boukouvala S (2008) Arylamine N-acetyltransferases: from structure to function. Drug Metab Rev 40: 479-510.
    • 4. Hein DW (2009) N-acetyltransferase SNPs: emerging concepts serve as a paradigm for understanding complexities of personalized medicine. Expert Opin Drug Metab Toxicol 5: 353-366.
    • 5. Sinclair JC, Sandy J, Delgoda R, Sim E, Noble ME (2000) Structure of arylamine N-acetyltransferase reveals a catalytic triad. Nat Struct Biol 7: 560- 564.
    • 6. Wu H, Dombrovsky L, Tempel W, Martin F, Loppnau P, et al. (2007) Structural basis of substrate-binding specificity of human arylamine N-acetyltransferases. J Biol Chem 282: 30189-30197.
    • 7. Goodfellow GH, Dupret JM, Grant DM (2000) Identification of amino acids imparting acceptor substrate selectivity to human arylamine acetyltransferases NAT1 and NAT2. Biochem J 348: 159-166.
    • 8. Sinclair J, Sim E (1997) A fragment consisting of the first 204 amino-terminal amino acids of human arylamine N-acetyltransferase one (NAT1) and the first transacetylation step of catalysis. Biochem Pharmacol 53: 11-16.
    • 9. Derewlany LO, Knie B, Koren G (1994) Arylamine N-acetyltransferase activity of the human placenta. J Pharmacol Exp Ther 269: 756-760.
    • 10. Derewlany LO, Knie B, Koren G (1994) Human placental transfer and metabolism of p-aminobenzoic acid. J Pharmacol Exp Ther 269: 761-765.
    • 11. Smelt VA, Upton A, Adjaye J, Payton MA, Boukouvala S, et al. (2000) Expression of arylamine N-acetyltransferases in pre-term placentas and in human pre-implantation embryos. Hum Mol Genet 9: 1101-1107.
    • 12. Pacifici GM, Bencini C, Rane A (1986) Acetyltransferase in humans: development and tissue distribution. Pharmacology 32: 283-291.
    • 13. Minchin RF (1995) Acetylation of p-aminobenzoylglutamate, a folic acid catabolite, by recombinant human arylamine N-acetyltransferase and U937 cells. Biochem J 307: 1-3.
    • 14. Ward A, Summers MJ, Sim E (1995) Purification of recombinant human Nacetyltransferase type 1 (NAT1) expressed in E. coli and characterization of its potential role in folate metabolism. Biochem Pharmacol 49: 1759-1767.
    • 15. Minchin RF, Hanna PE, Dupret JM, Wagner CR, Rodrigues-Lima F, et al. (2007) Arylamine N-acetyltransferase I. Int J Biochem Cell Biol 39: 1999-2005.
    • 16. Murphy M, Seagroatt V, Hey K, O'Donnell M, Godden M, et al. (1996) Neural tube defects 1974-94-down but not out. Arch Dis Child Fetal Neonatal Ed 75: F133-134.
    • 17. Jensen LE, Hoess K, Mitchell LE, Whitehead AS (2006) Loss of function polymorphisms in NAT1 protect against spina bifida. Hum Genet 120: 52-57.
    • 18. Erickson RP (2010) Genes, environment, and orofacial clefting: N-acetyltransferase and folic acid. J Craniofac Surg 21: 1384-1387.
    • 19. Czeizel AE, Timar L, Sarkozi A (1999) Dose-dependent effect of folic acid on the prevention of orofacial clefts. Pediatrics 104: e66.
    • 20. Kawamura A, Westwood I, Wakefield L, Long H, Zhang N, et al. (2008) Mouse N-acetyltransferase type 2, the homologue of human N-acetyltransferase type 1. Biochem Pharmacol 75: 1550-1560.
    • 21. Boukouvala S, Price N, Sim E (2002) Identification and functional characterization of novel polymorphisms associated with the genes for arylamine Nacetyltransferases in mice. Pharmacogenetics 12: 385-394.
    • 22. Martell KJ, Levy GN, Weber WW (1992) Cloned mouse N-acetyltransferases: enzymatic properties of expressed Nat-1 and Nat-2 gene products. Mol Pharmacol 42: 265-272.
    • 23. Hein DW, Boukouvala S, Grant DM, Minchin RF, Sim E (2008) Changes in consensus arylamine N-acetyltransferase gene nomenclature. Pharmacogenet Genomics 18: 367-368.
    • 24. Mitchell MK, Futscher BW, McQueen CA (1999) Developmental expression of N-acetyltransferases in C57BI/6 mice. Drug Metab Dispos 27: 261-264.
    • 25. Stanley LA, Copp AJ, Pope J, Rolls S, Smelt V, et al. (1998) Immunochemical detection of arylamine N-acetyltransferase during mouse embryonic development and in adult mouse brain. Teratology 58: 174-182.
    • 26. Ward A, Hickman D, Gordon JW, Sim E (1992) Arylamine N-acetyltransferase in human red blood cells. Biochem Pharmacol 44: 1099-1104.
    • 27. Payton M, Smelt V, Upton A, Sim E (1999) A method for genotyping murine arylamine N-acetyltransferase type 2 (NAT2): a gene expressed in preimplantation embryonic stem cells encoding an enzyme acetylating the folate catabolite p-aminobenzoylglutamate. Biochem Pharmacol 58: 779-785.
    • 28. Cornish VA, Pinter K, Boukouvala S, Johnson N, Labrousse C, et al. (2003) Generation and analysis of mice with a targeted disruption of the arylamine Nacetyltransferase Type 2 gene. Pharmacogenomics J 3: 169-177.
    • 29. Wakefield L, Cornish V, Long H, Griffiths WJ, Sim E (2007) Deletion of a xenobiotic metabolizing gene in mice affects folate metabolism. Biochem Biophys Res Commun 364: 556-560.
    • 30. Witham KL, Butcher NJ, Sugamori KS, Brenneman D, Grant DM, et al. (2013) 5-Methyl-tetrahydrofolate and the S-adenosylmethionine cycle in C57BL/6J Mouse Tissues: Gender differences and effects of arylamine N-acetyltransferase1 deletion. PLoS One 8: e77923.
    • 31. Wakefield L, Long H, Lack N, Sim E (2007) Ocular defects associated with a null mutation in the mouse arylamine N-acetyltransferase 2 gene. Mamm Genome 18: 270-276.
    • 32. Cao W, Chau B, Hunter R, Strnatka D, McQueen CA, et al. (2005) Only low levels of exogenous N-acetyltransferase can be achieved in transgenic mice. Pharmacogenomics J 5: 255-261.
    • 33. Erickson RP, Cao W, Acuna DK, Strnatka DW, Hunter RJ, et al. (2008) Confirmation of the role of N-acetyltransferase 2 in teratogen-induced cleft palate using transgenics and knockouts. Mol Reprod Dev 75: 1071-1076.
    • 34. Sim E, Pinter K, Mushtaq A, Upton A, Sandy J, et al. (2003) Arylamine Nacetyltransferases: a pharmacogenomic approach to drug metabolism and endogenous function. Biochem Soc Trans 31: 615-619.
    • 35. Sim E, Westwood I, Fullam E (2007) Arylamine N-acetyltransferases. Expert Opin Drug Metab Toxicol 3: 169-184.
    • 36. Wakefield L, Boukouvala S, Sim E (2010) Characterisation of CpG methylation in the upstream control region of mouse Nat2: evidence for a gene-environment interaction in a polymorphic gene implicated in folate metabolism. Gene 452: 16-21.
    • 37. Kim SJ, Kang HS, Chang HL, Jung YC, Sim HB, et al. (2008) Promoter hypomethylation of the N-acetyltransferase 1 gene in breast cancer. Oncol Rep 19: 663-668.
    • 38. Adam PJ, Berry J, Loader JA, Tyson KL, Craggs G, et al. (2003) Arylamine Nacetyltransferase-1 is highly expressed in breast cancers and conveys enhanced growth and resistance to etoposide in vitro. Mol Cancer Res 1: 826-835.
    • 39. Johansson I, Nilsson C, Berglund P, Lauss M, Ringner M, et al. (2012) Gene expression profiling of primary male breast cancers reveals two unique subgroups and identifies N-acetyltransferase-1 (NAT1) as a novel prognostic biomarker. Breast Cancer Res 14: R31.
    • 40. Wakefield L, Robinson J, Long H, Ibbitt JC, Cooke S, et al. (2008) Arylamine Nacetyltransferase 1 expression in breast cancer cell lines: a potential marker in estrogen receptor-positive tumors. Genes Chromosomes Cancer 47: 118-126.
    • 41. Russell AJ, Westwood IM, Crawford MH, Robinson J, Kawamura A, et al. (2009) Selective small molecule inhibitors of the potential breast cancer marker, human arylamine N-acetyltransferase 1, and its murine homologue, mouse arylamine N-acetyltransferase 2. Bioorg Med Chem 17: 905-918.
    • 42. Ballester PJ, Westwood I, Laurieri N, Sim E, Richards WG (2010) Prospective virtual screening with Ultrafast Shape Recognition: the identification of novel inhibitors of arylamine N-acetyltransferases. J R Soc Interface 7: 335-342.
    • 43. Tiang JM, Butcher NJ, Minchin RF (2010) Small molecule inhibition of arylamine N-acetyltransferase Type I inhibits proliferation and invasiveness of MDA-MB-231 breast cancer cells. Biochem Biophys Res Commun 393: 95- 100.
    • 44. Laurieri N, Crawford MH, Kawamura A, Westwood IM, Robinson J, et al. (2010) Small molecule colorimetric probes for specific detection of human arylamine N-acetyltransferase 1, a potential breast cancer biomarker. J Am Chem Soc 132: 3238-3239.
    • 45. Laurieri N, Egleton JE, Varney A, Thinnes CC, Quevedo CE, et al. (2013) A novel color change mechanism for breast cancer biomarker detection: naphthoquinones as specific ligands of human arylamine N-acetyltransferase 1. PLoS One 8: e70600.
    • 46. Dairou J, Atmane N, Dupret JM, Rodrigues-Lima F (2003) Reversible inhibition of the human xenobiotic-metabolizing enzyme arylamine N-acetyltransferase 1 by S-nitrosothiols. Biochem Biophys Res Commun 307: 1059-1065.
    • 47. Westwood IM, Holton SJ, Rodrigues-Lima F, Dupret JM, Bhakta S, et al. (2005) Expression, purification, characterization and structure of Pseudomonas aeruginosa arylamine N-acetyltransferase. Biochem J 385: 605-612.
    • 48. Sandy J, Mushtaq A, Kawamura A, Sinclair J, Sim E, et al. (2002) The structure of arylamine N-acetyltransferase from Mycobacterium smegmatis-an enzyme which inactivates the anti-tubercular drug, isoniazid. J Mol Biol 318: 1071-1083.
    • 49. Fullam E, Kawamura A, Wilkinson H, Abuhammad A, Westwood I, et al. (2009) Comparison of the arylamine N-acetyltransferase from Mycobacterium marinum and Mycobacterium tuberculosis. Protein J 28: 281-293.
    • 50. Abuhammad A, Lack N, Schweichler J, Staunton D, Sim RB, et al. (2011) Improvement of the expression and purification of Mycobacterium tuberculosis arylamine N-acetyltransferase (TBNAT) a potential target for novel antitubercular agents. Protein Expr Purif.
    • 51. Sandy J, Holton S, Fullam E, Sim E, Noble M (2005) Binding of the antitubercular drug isoniazid to the arylamine N-acetyltransferase protein from Mycobacterium smegmatis. Protein Sci 14: 775-782.
    • 52. Fullam E, Westwood IM, Anderton MC, Lowe ED, Sim E, et al. (2008) Divergence of cofactor recognition across evolution: Coenzyme A binding in a prokaryotic arylamine N-acetyltransferase. J Mol Biol 375: 178-191.
    • 53. Wang H, Vath GM, Kawamura A, Bates CA, Sim E, et al. (2005) Overexpression, purification, and characterization of recombinant human arylamine N-acetyltransferase 1. Protein J 24: 65-77.
    • 54. Grant DM, Blum M, Beer M, Meyer UA (1991) Monomorphic and polymorphic human arylamine N-acetyltransferases: a comparison of liver isozymes and expressed products of two cloned genes. Mol Pharmacol 39: 184- 191.
    • 55. Brooke EW, Davies SG, Mulvaney AW, Pompeo F, Sim E, et al. (2003) An approach to identifying novel substrates of bacterial arylamine N-acetyltransferases. Bioorg Med Chem 11: 1227-1234.
    • 56. Andres HH, Klem AJ, Szabo SM, Weber WW (1985) New spectrophotometric and radiochemical assays for acetyl-CoA: arylamine N-acetyltransferase applicable to a variety of arylamines. Anal Biochem 145: 367-375.
    • 57. Westwood IM, Kawamura A, Fullam E, Russell AJ, Davies SG, et al. (2006) Structure and mechanism of arylamine N-acetyltransferases. Curr Top Med Chem 6: 1641-1654.
    • 58. Patel SS, Walt DR (1987) Substrate specificity of acetyl Coenzyme A synthetase. J Biol Chem 262: 7132-7134.
    • 59. Wu WJ, Tonge PJ, Raleigh DP (1998) Stereospecific H-1 and C-13 NMR assignments of crotonyl CoA and hexadienoyl CoA: Conformational analysis and comparison with protein-CoA complexes. Journal of the American Chemical Society 120: 9988-9994.
    • 60. Rossi C, Donati A, Sansoni MR (1993) Folic-Acid - Solution Structure and NMR Strategy for Conformational-Analysis. Spectrosc Lett 26: 1603-1611.
    • 61. DeLano WL (2002) PyMOL: An open-source molecular graphics tool. DeLano Scientific, San Carlos, California, USA; http://www.ccp4.ac.uk/newsletters/ newsletter40/11_pymol.pdf.
    • 62. Verdonk ML, Cole JC, Hartshorn MJ, Murray CW, Taylor RD (2003) Improved protein-ligand docking using GOLD. Proteins 52: 609-623.
    • 63. Mushtaq A, Payton M, Sim E (2002) The COOH terminus of arylamine Nacetyltransferase from Salmonella typhimurium controls enzymic activity. J Biol Chem 277: 12175-12181.
    • 64. Wang H, Liu L, Hanna PE, Wagner CR (2005) Catalytic mechanism of hamster arylamine N-acetyltransferase 2. Biochemistry 44: 11295-11306.
    • 65. Wang H, Vath GM, Gleason KJ, Hanna PE, Wagner CR (2004) Probing the mechanism of hamster arylamine N-acetyltransferase 2 acetylation by active site modification, site-directed mutagenesis, and pre-steady state and steady state kinetic studies. Biochemistry 43: 8234-8246.
    • 66. Jencks WP, Gresser M, Valenzuela MS, Huneeus FC (1972) Acetyl Coenzyme A: arylamine acetyltransferase. Measurement of the steady state concentration of the acetyl-enzyme intermediate. J Biol Chem 247: 3756-3760.
    • 67. Claridge TDW, Davies SG, Polywka MEC, Roberts PM, Russell AJ, et al. (2008) ''Pure by NMR''? Organic Letters 10: 5433-5436.
    • 68. Abuhammad AM, Lowe ED, Fullam E, Noble M, Garman EF, et al. (2010) Probing the architecture of the Mycobacterium marinum arylamine N-acetyltransferase active site. Protein Cell 1: 384-392.
    • 69. Matte C, Mackedanz V, Stefanello FM, Scherer EB, Andreazza AC, et al. (2009) Chronic hyperhomocysteinemia alters antioxidant defenses and increases DNA damage in brain and blood of rats: protective effect of folic acid. Neurochem Int 54: 7-13.
    • 70. Moat SJ, Bonham JR, Cragg RA, Powers HJ (2000) Elevated plasma homocysteine elicits an increase in antioxidant enzyme activity. Free Radic Res 32: 171-179.
    • 71. Egleton JE, Thinnes CC, Seden PT, Laurieri N, Lee SP, et al. (2014) Structureactivity relationships and colorimetric properties of specific probes for the putative cancer biomarker human arylamine N-acetyltransferase 1. Bioorg Med Chem In press.
    • 72. Lam Y-F, Kotowycz G (1972) Self association of folic acid in aqueous solution by Proton Magnetic Resonance. Canadian Journal of Chemistry 50: 2357-2363.
    • 73. Rossi C, Donati A, Sansoni MR (1993) Folic acid: Solution structure and NMR strategy for conformational analysis. Spectrosc Lett 26: 1603-1611.
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