A new H/D exchange- and mass spectrometry-based method for thermodynamic analysis of protein-DNA interactions.

Journal Article

The application of SUPREX (stability of unpurified proteins from rates of H/D exchange) to the thermodynamic analysis of protein-DNA complexes is described. A series of five model protein-DNA complexes involving two known DNA binding proteins, Arc repressor and CopG, were analyzed in order to determine the accuracy, precision, and generality of the SUPREX technique for quantifying the strength of protein-DNA interactions. For protein-DNA complexes that reversibly unfold in a two-state manner, we demonstrate that reasonably precise Kd values in agreement with those determined by conventional techniques can be determined by SUPREX. In the case of protein-DNA complexes that are not well modeled by a two-state unfolding mechanism, we find that relative binding affinities can be determined in the SUPREX experiment.

Full Text

Duke Authors

Cited Authors

  • Ma, L; Fitzgerald, MC

Published Date

  • December 2003

Published In

Volume / Issue

  • 10 / 12

Start / End Page

  • 1205 - 1213

PubMed ID

  • 14700628

International Standard Serial Number (ISSN)

  • 1074-5521

Language

  • eng

Conference Location

  • United States