Rapid characterization of folding and binding interactions with thermolabile ligands by DSC

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Date

2016-10-23

Authors

Harkness V, Robert W
Slavkovic, Sladjana
Johnson, Philip E.
Mittermaier, Anthony K.

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Publisher

The Royal Society of Chemistry

Abstract

Differential scanning calorimetry (DSC) is a powerful technique for measuring tight biomolecular interactions. However, many pharma- ceutically relevant ligands are chemically unstable at the high temperatures used in DSC analyses. Thus, measuring binding inter- actions is challenging because the concentrations of ligands and thermally-converted products are constantly changing within the calorimeter cell. Using experimental data for two DNA aptamers that bind to the thermolabile ligand cocaine, we present a new global fitting analysis that yields the complete set of folding and binding parameters for the initial and final forms of the ligand from a pair of DSC experiments, while accounting for the thermal conversion. Furthermore, we show that the rate constant for thermolabile ligand conversion may be obtained with only one additional DSC dataset.

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Citation

Chem. Commun. 52 (2016): 13471--13474