Isolation and partial characterization of the adduct formed by 13-hydroxyellipticine with deoxyguanosine in DNA
Language English Country Sweden Media print
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
18987592
PII: NEL290508A19
Knihovny.cz E-resources
- MeSH
- Acetyl Coenzyme A metabolism MeSH
- DNA Adducts chemistry isolation & purification MeSH
- Arylamine N-Acetyltransferase metabolism MeSH
- Arylsulfotransferase metabolism MeSH
- Autoradiography MeSH
- Deoxyguanosine chemistry MeSH
- DNA chemistry MeSH
- Ellipticines chemistry MeSH
- Phosphoadenosine Phosphosulfate chemistry MeSH
- Indicators and Reagents MeSH
- Isoenzymes metabolism MeSH
- Catalysis MeSH
- Hydrogen-Ion Concentration MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- 13-hydroxyellipticine MeSH Browser
- Acetyl Coenzyme A MeSH
- DNA Adducts MeSH
- Arylamine N-Acetyltransferase MeSH
- Arylsulfotransferase MeSH
- Deoxyguanosine MeSH
- DNA MeSH
- Ellipticines MeSH
- Phosphoadenosine Phosphosulfate MeSH
- Indicators and Reagents MeSH
- Isoenzymes MeSH
- N-acetyltransferase 1 MeSH Browser
- NAT2 protein, human MeSH Browser
- SULT1A1 protein, human MeSH Browser
- SULT1A2 protein, human MeSH Browser
OBJECTIVES: Ellipticine is a potent antineoplastic agent exhibiting multiple mechanisms of its action. Recently, we have found that 13-hydroxyellipticine, formed from ellipticine as the predominant metabolite in human livers, is bound to deoxyguanosine in DNA, generating the major DNA adduct in vivo and in vitro. The development of the methods suitable for the preparation of this adduct in the amounts sufficient for identification of its structure and those for its isolation and partial characterization is the aim of this study. METHODS: High performance liquid chromatography (HPLC) was employed for separation of 13-hydroxyellipticine-mediated deoxyguanosine adduct. The 32P-postlabeling technique was utilized to detect this adduct in DNA. RESULTS: The formation of the 13-hydroxyellipticine-derived deoxyguanosine adduct in DNA in vitro was increased under the alkaline pH of the incubations and by the formation of the sulfate and acetate conjugates of 13-hydroxyellipticine generated by reactions with 3'-phosphoadenosine-5'-phosphosulfate (PAPS) or acetyl-coenzyme A (acetyl-CoA) catalyzed by human sulfotransferases (SULTs) 1A1 and 1A2 and N,O-acetyltransferases (NATs) 1 and 2. The HPLC method suitable for separation the 13-hydroxyellipticine-derived deoxyguanosine adduct from other reactants, deoxyguanosine and 13-hydroxyellipticine, was developed. The structure of this adduct is proposed to correspond to the product formed from ellipticine-13-ylium with the exocyclic 2-NH2 group of guanine in DNA. CONCLUSIONS: The data are the first report on HPLC isolation of the deoxyguanosine adduct formed by 13-hydroxyellipticine in DNA and its partial characterization.
Formation of DNA adducts by ellipticine and its micellar form in rats - a comparative study
Ellipticine cytotoxicity to cancer cell lines - a comparative study