title: 2-Amino-9H-pyrido[2,3-b]indole (AαC) Adducts and Thiol Oxidation of Serum Albumin as Potential Biomarkers of Tobacco Smoke.
abstract: 2-Amino-9H-pyrido[2,3-b]indole (AαC) is a carcinogenic heterocyclic aromatic amine formed during the combustion of tobacco. AαC undergoes bioactivation to form electrophilic N-oxidized metabolites that react with DNA to form adducts, which can lead to mutations. Many genotoxicants and toxic electrophiles react with human serum albumin (albumin); however, the chemistry of reactivity of AαC with proteins has not been studied. The genotoxic metabolites, 2-hydroxyamino-9H-pyrido[2,3-b]indole (HONH-AαC), 2-nitroso-9H-pyrido[2,3-b]indole (NO-AαC), N-acetyloxy-2-amino-9H-pyrido[2,3-b]indole (N-acetoxy-AαC), and their [(13)C6]AαC-labeled homologues were reacted with albumin. Sites of adduction of AαC to albumin were identified by data-dependent scanning and targeted bottom-up proteomics approaches employing ion trap and Orbitrap MS. AαC-albumin adducts were formed at Cys(34), Tyr(140), and Tyr(150) residues when albumin was reacted with HONH-AαC or NO-AαC. Sulfenamide, sulfinamide, and sulfonamide adduct formation occurred at Cys(34) (AαC-Cys(34)). N-Acetoxy-AαC also formed an adduct at Tyr(332). Albumin-AαC adducts were characterized in human plasma treated with N-oxidized metabolites of AαC and human hepatocytes exposed to AαC. High levels of N-(deoxyguanosin-8-yl)-AαC (dG-C8-AαC) DNA adducts were formed in hepatocytes. The Cys(34) was the sole amino acid of albumin to form adducts with AαC. Albumin also served as an antioxidant and scavenged reactive oxygen species generated by metabolites of AαC in hepatocytes; there was a strong decrease in reduced Cys(34), whereas the levels of Cys(34) sulfinic acid (Cys-SO2H), Cys(34)-sulfonic acid (Cys-SO3H), and Met(329) sulfoxide were greatly increased. Cys(34) adduction products and Cys-SO2H, Cys-SO3H, and Met(329) sulfoxide may be potential biomarkers to assess exposure and oxidative stress associated with AαC and other arylamine toxicants present in tobacco smoke.
authors.affiliation: From the Masonic Cancer Center and Department of Medicinal Chemistry, University of Minnesota, Minneapolis, Minnesota 55455 and.
authors.forename: Khyatiben V
authors.initials: KV
authors.lastname: Pathak
authors.affiliation: UMR INSERM 1085 IRSET, Rennes 1 University, UMS 3480 Biosit, F-35043 Rennes, France.
authors.forename: Medjda
authors.initials: M
authors.lastname: Bellamri
authors.affiliation: From the Masonic Cancer Center and Department of Medicinal Chemistry, University of Minnesota, Minneapolis, Minnesota 55455 and.
authors.forename: Yi
authors.initials: Y
authors.lastname: Wang
authors.affiliation: UMR INSERM 1085 IRSET, Rennes 1 University, UMS 3480 Biosit, F-35043 Rennes, France.
authors.forename: Sophie
authors.initials: S
authors.lastname: Langouët
authors.affiliation: From the Masonic Cancer Center and Department of Medicinal Chemistry, University of Minnesota, Minneapolis, Minnesota 55455 and rturesky@umn.edu.
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Turesky
mesh_terms: D020816:Amino Acid Motifs; D015415:Biomarkers / Q000737:chemistry / Q000378:metabolism; D002243:Carbolines / Q000009:adverse effects / Q000737:chemistry* / Q000378:metabolism; D018736:DNA Adducts / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D022781:Hepatocytes / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D010084:Oxidation-Reduction; D012709:Serum Albumin / Q000737:chemistry* / Q000378:metabolism; D012906:Smoke / Q000009:adverse effects*; D013438:Sulfhydryl Compounds / Q000737:chemistry* / Q000378:metabolism; D062789:Tobacco Products
chemical_list: D015415:Biomarkers; D002243:Carbolines; D018736:DNA Adducts; D012709:Serum Albumin; D012906:Smoke; D013438:Sulfhydryl Compounds; C017822:2-amino-9H-pyrido(2,3-b)indole
keywords: Carcinogenesis; Cigarette Smoke; DNA Damage; Oxidative Stress; Protein Chemical Modification