Entropically-driven binding of mithramycin in the minor groove of C/G-rich DNA sequences.
The antitumour antibiotic mithramycin A (MTA) is a DNA minor-groove binding ligand. It binds to C/G-rich tracts as a dimer that forms in the presence of divalent cations such as Mg(2+). Differential scanning calorimetry, UV thermal denaturation, isothermal titration calorimetry and competition dialysis were used, together with computations of the hydrophobic free energy of binding, to determine the thermodynamic profile of MTA binding to DNA. The results were compared to those obtained in parallel using the structurally related mithramycin SK (MSK). The binding of MTA to salmon testes DNA determined by UV melting studies (K(obs) = 1.2 (+/-0.3) x 10(5) M(-1)) is tighter than that of MSK (2.9 (+/-1.0) x 10(4) M(-1)) at 25 degrees C. Competition dialysis studies showed a tighter MTA binding to both salmon testes DNA (42% C + G) and Micrococcus lysodeikticus DNA (72% C + G). The thermodynamic analysis of binding data at 25 degrees C shows that the binding of MTA and MSK to DNA is entropically driven, dominated by the hydrophobic transfer of the antibiotics from solution to the DNA-binding site. Direct molecular recognition between MTA or MSK and DNA through hydrogen bonding and van der Waals contacts may also contribute significantly to complex formation.
Barceló
Francisca
F
Departament de Biologia Fundamental i Ciencies de la Salut, Universitat de les Illes Balears, Palma de Mallorca, Spain.
Scotta
Claudia
C
Ortiz-Lombardía
Miguel
M
Méndez
Carmen
C
Salas
José A
JA
Portugal
José
J
D000903:Antibiotics, Antineoplastic / Q000737:chemistry*; D002151:Calorimetry; D004247:DNA / Q000737:chemistry*; D003956:Dialysis; D019277:Entropy; D020862:GC Rich Sequence; D006358:Hot Temperature; D008958:Models, Molecular; D009690:Nucleic Acid Conformation; D009691:Nucleic Acid Denaturation; D008926:Plicamycin / Q000031:analogs & derivatives* / Q000737:chemistry; D013056:Spectrophotometry, Ultraviolet
D000903:Antibiotics, Antineoplastic; C520007:mithramycin SK; D004247:DNA; C066851:mithramycin A; D008926:Plicamycin