Abstract
We 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 K(D) 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 (k(off): 0.02 s(-1)) and FABP (k(off): 0.01 s(-1)) lesions than to FAF (k(off): 0.04 s(-1)).
Publication types
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Research Support, N.I.H., Extramural
MeSH terms
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2-Acetylaminofluorene / chemistry*
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Aminobiphenyl Compounds / chemistry*
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DNA / chemistry*
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DNA / metabolism
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DNA Adducts / chemistry
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DNA Polymerase I / metabolism*
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Deoxyadenine Nucleotides / chemistry
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Deoxycytosine Nucleotides / chemistry
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Deoxyguanine Nucleotides / chemistry
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Fluorenes / chemistry*
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Kinetics
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Surface Plasmon Resonance
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Thymine Nucleotides / chemistry
Substances
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Aminobiphenyl Compounds
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DNA Adducts
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Deoxyadenine Nucleotides
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Deoxycytosine Nucleotides
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Deoxyguanine Nucleotides
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Fluorenes
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Thymine Nucleotides
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4-biphenylamine
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2'-deoxycytidine 5'-triphosphate
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2-aminofluorene
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deoxyguanosine triphosphate
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DNA
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2-Acetylaminofluorene
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DNA Polymerase I
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2'-deoxyadenosine triphosphate
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thymidine 5'-triphosphate