title: 7-glutathione pyrrole adduct: a potential DNA reactive metabolite of pyrrolizidine alkaloids.
abstract: Pyrrolizidine alkaloid (PA)-containing plants are the most common poisonous plants affecting livestock, wildlife, and humans. PAs require metabolic activation to form pyrrolic metabolites to exert cytotoxicity and tumorigenicity. We previously determined that metabolism of tumorigenic PAs produced four DNA adducts, designated as DHP-dG-3, DHP-dG-4, DHP-dA-3, and DHP-dA-4, that are responsible for liver tumor initiation. 7-Glutathione-(±)-6,7-dihydro-1-hydroxymethyl-5H-pyrrolizine (7-GS-DHP), formed in vivo and in vitro, and 7,9-di-GS-DHP, formed in vitro, are both considered detoxified metabolites. However, in this study we determined that incubation of 7-GS-DHP with 2'-deoxyguanosine (dG) and 2'-deoxyadenosine (dA) yields DHP-dG-3, DHP-dG-4, DHP-dA-3, and DHP-dA-4 adducts as well as the reactive metabolite DHP. Furthermore, reaction of 7-GS-DHP with calf thymus DNA in aqueous solution at 37 °C for 4, 8, 16, 24, 48, or 72 h, followed by enzymatic hydrolysis yielded DHP-dG-3, DHP-dG-4, DHP-dA-3, and DHP-dA-4 adducts. Under our current experimental conditions, DHP-dA-3 and DHP-dA-4 adducts were formed in a trace amount from the reaction of 7,9-di-GS-DHP with dA. No DHP-dG-3 or DHP-dG-4 adducts were detected from the reaction of 7,9-di-GS-DHP with dG. This study represents the first report that the 7-GS-DHP adduct can be a potential reactive metabolite of PAs leading to DNA adduct formation.
authors.affiliation: †National Center for Toxicological Research, U.S. Food and Drug Administration, Jefferson, Arkansas 72079, United States.
authors.forename: Qingsu
authors.initials: Q
authors.lastname: Xia
authors.affiliation: †National Center for Toxicological Research, U.S. Food and Drug Administration, Jefferson, Arkansas 72079, United States.
authors.forename: Liang
authors.initials: L
authors.lastname: Ma
authors.affiliation: †National Center for Toxicological Research, U.S. Food and Drug Administration, Jefferson, Arkansas 72079, United States.
authors.forename: Xiaobo
authors.initials: X
authors.lastname: He
authors.affiliation: ‡Biotranex LLC, Monmouth Junction, New Jersey 08852, United States.
authors.forename: Lining
authors.initials: L
authors.lastname: Cai
authors.affiliation: †National Center for Toxicological Research, U.S. Food and Drug Administration, Jefferson, Arkansas 72079, United States.
authors.forename: Peter P
authors.initials: PP
authors.lastname: Fu
mesh_terms: D000818:Animals; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D018736:DNA Adducts / Q000737:chemistry*; D005978:Glutathione / Q000737:chemistry*; D011758:Pyrroles / Q000737:chemistry*; D011763:Pyrrolizidine Alkaloids / Q000737:chemistry*
chemical_list: D018736:DNA Adducts; D011758:Pyrroles; D011763:Pyrrolizidine Alkaloids; D005978:Glutathione