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Binary and Ternary Binding Affinities between Exonuclease-Deficient Klenow Fragment (Kf-exo–) and Various Arylamine DNA Lesions Characterized by Surface Plasmon Resonance
journal contribution
posted on 2012-08-20, 00:00 authored by V. G. Vaidyanathan, Lifang Xu, Bongsup P. ChoWe used surface plasmon resonance (SPR) to characterize
the binding
interactions between the exonulease-free Klenow fragment (Kf-exo–) 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 KD values in the order of dCTP ≪ dTTP < dATP
< dGTP. In contrast, minimal selectivity was observed for the modified
templates with Kf-exo– binding tighter to the FAAF
(koff: 0.02 s–1) and
FABP (koff: 0.01 s–1) lesions than to FAF (koff: 0.04 s–1).