Enthalpy/entropy compensation: influence of DNA flanking sequence on the binding of 7-amino actinomycin D to its primary binding site in short DNA duplexes.
The effect of the context of the flanking sequence on ligand binding to DNA oligonucleotides that contain consensus binding sites was investigated for the binding of the intercalator 7-amino actinomycin D. Seven self-complementary DNA oligomers each containing a centrally located primary binding site, 5'-A-G-C-T-3', flanked on either side by the sequences (AT)(n) or (AA)(n) (with n = 2, 3, 4) and AA(AT)(2), were studied. For different flanking sequences, (AA)(n)-series or (AT)(n)-series, differential fluorescence enhancements of the ligand due to binding were observed. Thermodynamic studies indicated that the flanking sequences not only affected DNA stability and secondary structure but also modulated ligand binding to the primary binding site. The magnitude of the ligand binding affinity to the primary site was inversely related to the sequence dependent stability. The enthalpy of ligand binding was directly measured by isothermal titration calorimetry, and this made it possible to parse the binding free energy into its energetic and entropic terms. Our results reveal a pronounced enthalpy-entropy compensation for 7-amino actinomycin D binding to this family of oligonucleotides and suggest that the DNA sequences flanking the primary binding site can strongly influence ligand recognition of specific sites on target DNA molecules.
Qu
Xiaogang
X
Key Laboratory of Rare Earth Chemistry and Physics, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, Jilin 130022, China. xqu@ciac.jl.cn
Ren
Jinsong
J
Riccelli
Peter V
PV
Benight
Albert S
AS
Chaires
Jonathan B
JB
D000970:Antineoplastic Agents / Q000737:chemistry; D001483:Base Sequence; D001665:Binding Sites; D002151:Calorimetry; D002942:Circular Dichroism; D004247:DNA / Q000737:chemistry*; D003609:Dactinomycin / Q000031:analogs & derivatives* / Q000737:chemistry*; D019277:Entropy; D006358:Hot Temperature; D007364:Intercalating Agents / Q000737:chemistry*; D008024:Ligands; D009692:Nucleic Acid Heteroduplexes / Q000737:chemistry*; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D013050:Spectrometry, Fluorescence
D000970:Antineoplastic Agents; D007364:Intercalating Agents; D008024:Ligands; D009692:Nucleic Acid Heteroduplexes; D009838:Oligodeoxyribonucleotides; D003609:Dactinomycin; C025942:7-aminoactinomycin D; D004247:DNA