A spectroscopic and calorimetric study of the melting behaviors of a "bent" and a "normal" DNA duplex: [d(GA4T4C)]2 versus [d(GT4A4C)]2.

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RESUMO

We have calorimetrically detected and energetically characterized a premelting event in the "bent" duplex [d(GA4T4C)]2 that is absent in the "normal" duplex [d(GT4A4C)]2. This premelting may correspond to the thermally induced "unbending" of the duplex. Specifically, we have used a combination of spectroscopic and calorimetric techniques to evaluate whether differences in the electrophoretic and hydrogen exchange properties of the bent duplex, [d(GA4T4C)]2, and the normal duplex, [d(GT4A4C)]2, are paralleled by differences in the equilibrium melting properties of these duplexes. Our results reveal that the bimolecular global meltings of both duplexes exhibit two-state behavior and are characterized by the expected thermodynamic changes, as well as the expected salt-dependencies. Significantly, however, at temperatures below duplex melting and over a similar temperature range in which the aberrant electrophoretic mobility is observed (approximately less than 35 degrees C), the bent duplex exhibits calorimetric premelting behavior absent in the normal duplex. Analysis of the calorimetric data in this preglobal melting domain (approximately less than 35 degrees C) allows us to estimate a lower-limit value of 1.6 kcal/(mol-base pair) for the thermally induced unbending of the decameric duplex.

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