title: Stoichiometric preference in copper-promoted oxidative DNA damage by ochratoxin A.
abstract: The ability of the fungal carcinogen, ochratoxin A (OTA, 1), to facilitate copper-promoted oxidative DNA damage has been assessed using supercoiled plasmid DNA (Form I)-agarose gel electrophoresis and gas chromatography-mass spectrometry with selected-ion monitoring (GC-MS-SIM). OTA is shown to promote oxidative cleavage of Form I DNA with optimal cleavage efficiency occurring under excess Cu(II) conditions. As the concentration of OTA was increased and present in excess of Cu(II) the cleavage was less effective. Parallel findings were found for the ability of the OTA-Cu mixture to facilitate oxidative base damage. Yields (lesions per 10(6) DNA bases) of modified bases upon exposure of calf-thymus DNA (CT-DNA) to OTA-H(2)O(2)-Cu(II) were diminished when the OTA:Cu ratio was increased to 5:1. Electrochemical studies carried out in methanol implicate a ligand-centered 2e oxidation of OTA in the presence of excess Cu(II), while product analyses utilizing electrospray mass spectrometry support the intermediacy of the quinone, OTQ (3), in Cu-promoted oxidation of OTA. The implications of these findings with regard to the mutagenicity of OTA are discussed.
authors.affiliation: Department of Chemistry, Wake Forest University, Winston-Salem, NC 27109-7486, USA. manderra@wfu.edu
authors.forename: Richard A
authors.initials: RA
authors.lastname: Manderville
authors.forename: M
authors.initials: M
authors.lastname: Wade Calcutt
authors.forename: Jian
authors.initials: J
authors.lastname: Dai
authors.forename: Gyungse
authors.initials: G
authors.lastname: Park
authors.forename: Ivan G
authors.initials: IG
authors.lastname: Gillman
authors.forename: Ronald E
authors.initials: RE
authors.lastname: Noftle
authors.forename: Abdul K
authors.initials: AK
authors.lastname: Mohammed
authors.forename: Mustafa
authors.initials: M
authors.lastname: Birincioglu
authors.forename: Miral
authors.initials: M
authors.lastname: Dizdaroglu
authors.forename: Henry
authors.initials: H
authors.lastname: Rodriguez
authors.forename: Steven A
authors.initials: SA
authors.lastname: Akman
mesh_terms: D000818:Animals; D016227:Benzoquinones / Q000737:chemistry; D002417:Cattle; D003300:Copper / Q000737:chemistry* / Q000633:toxicity; D004247:DNA / Q000737:chemistry; D004249:DNA Damage; D004278:DNA, Superhelical / Q000737:chemistry* / Q000187:drug effects; D004563:Electrochemistry / Q000379:methods; D004587:Electrophoresis, Agar Gel; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods; D006861:Hydrogen Peroxide / Q000737:chemistry; D009793:Ochratoxins / Q000737:chemistry* / Q000633:toxicity; D010084:Oxidation-Reduction; D010957:Plasmids / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization; D013056:Spectrophotometry, Ultraviolet / Q000379:methods
chemical_list: D016227:Benzoquinones; D004278:DNA, Superhelical; D009793:Ochratoxins; C025589:ochratoxin A; C004532:quinone; D003300:Copper; D004247:DNA; C427203:calf thymus DNA; D006861:Hydrogen Peroxide