Cargando…

Formation of DNA−Protein Cross-Links Between γ-Hydroxypropanodeoxyguanosine and EcoRI

[Image: see text] The toxicity of acrolein, an α,β-unsaturated aldehyde produced during lipid peroxidation, is attributable to its high reactivity toward DNA and cellular proteins. The major acrolein−DNA adduct, γ-hydroxypropano-2′-deoxyguanosine (γ-HOPdG), ring opens to form a reactive N(2)-oxoprop...

Descripción completa

Detalles Bibliográficos
Autores principales: VanderVeen, Laurie A., Harris, Thomas M., Jen-Jacobson, Linda, Marnett, Lawrence J.
Formato: Texto
Lenguaje:English
Publicado: American Chemical Society 2008
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2651693/
https://www.ncbi.nlm.nih.gov/pubmed/18690724
http://dx.doi.org/10.1021/tx800092g
Descripción
Sumario:[Image: see text] The toxicity of acrolein, an α,β-unsaturated aldehyde produced during lipid peroxidation, is attributable to its high reactivity toward DNA and cellular proteins. The major acrolein−DNA adduct, γ-hydroxypropano-2′-deoxyguanosine (γ-HOPdG), ring opens to form a reactive N(2)-oxopropyl moiety that cross-links to DNA and proteins. We demonstrate the ability of γ-HOPdG in a duplex oligonucleotide to cross-link to a protein (EcoRI) that specifically interacts with DNA at a unique sequence. The formation of a cross-link to EcoRI was dependent on the intimate binding of the enzyme to its γ-HOPdG-modified recognition site. Interestingly, the cross-link did not restrict the ability of EcoRI to cleave DNA substrates. However, stabilization of the cross-link by reduction of the Schiff base linkage resulted in loss of enzyme activity. This work indicates that the γ-HOPdG−EcoRI cross-link is in equilibrium with free oligonucleotide and enzyme. Reversal of cross-link formation allows EcoRI to effect enzymatic cleavage of competitor oligonucleotides.