Abstract

The changes in binding free energies due to cooperativity for oligodeoxyribonucleotides bound to adjacent sites by triple helix formation have been determined by quantitative affinity cleavage titrations. A 20-fold enhancement in equilibrium association constant is realized for an l lmer pyrimidine oligonucleotide binding in the presence of a neighboring bound site at 24 °C and pH 7.0 (25 mM Tris-OAc, 10 mM NaCl, 1 mM spermine). This corresponds to an increase in binding free energy of 1.8 ± 0.2 kcal mol^(-1). This cooperativity is not observed when the two binding sites are separated by one base pair.

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