10.1021/tx300289d.s001
V. G. Vaidyanathan
V. G.
Vaidyanathan
Lifang Xu
Lifang
Xu
Bongsup P. Cho
Bongsup P.
Cho
Binary and Ternary Binding
Affinities between Exonuclease-Deficient
Klenow Fragment (Kf-exo<sup>–</sup>) and Various Arylamine
DNA Lesions Characterized by Surface Plasmon Resonance
American Chemical Society
2012
surface plasmon resonance
KD values
Surface Plasmon ResonanceWe
dCTP
FABP
SPR
Various Arylamine DNA Lesions Characterized
dG adducts
FAF
arylamine carcinogens
Ternary Binding Affinities
koff
FAAF
Tight polymerase binding
Klenow
binding interactions
2012-08-20 00:00:00
Journal contribution
https://acs.figshare.com/articles/journal_contribution/Binary_and_Ternary_Binding_Affinities_between_Exonuclease_Deficient_Klenow_Fragment_Kf_exo_sup_sup_and_Various_Arylamine_DNA_Lesions_Characterized_by_Surface_Plasmon_Resonance/2495152
We used surface plasmon resonance (SPR) to characterize
the binding
interactions between the exonulease-free Klenow fragment (Kf-exo<sup>–</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 <i>K</i><sub>D</sub> values in the order of dCTP ≪ dTTP < dATP
< dGTP. In contrast, minimal selectivity was observed for the modified
templates with Kf-exo<sup>–</sup> binding tighter to the FAAF
(<i>k</i><sub>off</sub>: 0.02 s<sup>–1</sup>) and
FABP (<i>k</i><sub>off</sub>: 0.01 s<sup>–1</sup>) lesions than to FAF (<i>k</i><sub>off</sub>: 0.04 s<sup>–1</sup>).