SSB binding to ssDNA using isothermal titration calorimetry

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

9 Scopus citations

Abstract

Isothermal titration calorimetry (ITC) is a powerful method for studying protein-DNA interactions in solution. As long as binding is accompanied by an appreciable enthalpy change, ITC studies can yield quantitative information on stoichiometries, binding energetics (affinity, binding enthalpy and entropy) and potential site-site interactions (cooperativity). This can provide a full thermodynamic description of an interacting system which is necessary to understand the stability and specificity of protein-DNA interactions and to correlate the activities or functions of different species. Here we describe procedures to perform and analyze ITC studies using as examples, the E. coli SSB (homotetramer with 4 OB-folds) and D. radiodurans SSB (homodimer with 4 OB-folds). For oligomeric protein systems such as these, we emphasize the need to be aware of the likelihood that solution conditions will influence not only the affinity and enthalpy of binding but also the mode by which the SSB oligomer binds ssDNA.

Original languageEnglish
Title of host publicationSingle-Stranded DNA Binding Proteins
Subtitle of host publicationMethods and Protocols
PublisherHumana Press Inc.
Pages37-54
Number of pages18
ISBN (Print)9781627030311
DOIs
StatePublished - 2012

Publication series

NameMethods in Molecular Biology
Volume922
ISSN (Print)1064-3745

Keywords

  • DrSSB
  • EcoSSB
  • ITC
  • SSB-ssDNA thermodynamics
  • ssDNA binding

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