Binary and Ternary Binding Affinities between Exonuclease-Deficient Klenow Fragment (Kf-exo鈥?/sup>) and Various Arylamine DNA Lesions Characterized by Surface Plasmon Resonance

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• 作者：V. G. Vaidyanathan ; Lifang Xu ; Bongsup P. Cho
• 刊名：Chemical Research in Toxicology
• 出版年：2012
• 出版时间：August 20, 2012
• 年：2012
• 卷：25
• 期：8
• 页码：1568-1570
• 全文大小：217K
• 年卷期：v.25,no.8(August 20, 2012)
• ISSN：1520-5010

文摘

We used surface plasmon resonance (SPR) to characterize the binding interactions between the exonulease-free Klenow fragment (Kf-exo^{鈥?/sup>) and unmodified and modified dG adducts derived from arylamine carcinogens: fluorinated 2-aminofluorene (FAF), 2-acetylaminofluorene (FAAF), and 4-aminobiphenyl (FABP). Tight polymerase binding was detected with unmodified dG and the correct dCTP. The discrimination of correct versus incorrect nucleotides was pronounced with K_D values in the order of dCTP dTTP < dATP < dGTP. In contrast, minimal selectivity was observed for the modified templates with Kf-exo鈥?/sup> binding tighter to the FAAF (k_off: 0.02 s鈥?) and FABP (k_off: 0.01 s鈥?) lesions than to FAF (k_off: 0.04 s鈥?).}