Multi-wavelength analytical ultracentrifugation as a tool to characterise protein–DNA interactions in solution

Christopher R. Horne, Amy Henrickson, Borries Demeler, Renwick C.J. Dobson

Research output: Contribution to journalArticlepeer-review

10 Scopus citations


Understanding how proteins interact with DNA, and particularly the stoichiometry of a protein–DNA complex, is key information needed to elucidate the biological role of the interaction, e.g. transcriptional regulation. Here, we present an emerging analytical ultracentrifugation method that features multi-wavelength detection to characterise complex mixtures by deconvoluting the spectral signals of the interaction partners into separate sedimentation profiles. The spectral information obtained in this experiment provides direct access to the molar stoichiometry of the interacting system to complement traditional hydrodynamic information. We demonstrate this approach by characterising a multimeric assembly process between the transcriptional repressor of bacterial sialic acid metabolism, NanR and its DNA-binding sequence. The method introduced in this study can be extended to quantitatively analyse any complex interaction in solution, providing the interaction partners have different optical properties.

Original languageEnglish
Pages (from-to)819-827
Number of pages9
JournalEuropean Biophysics Journal
Issue number8
StatePublished - Dec 2020


  • Multi-wavelength analytical ultracentrifugation
  • NanR
  • Protein–DNA interaction
  • Sedimentation velocity
  • Sialic acid


Dive into the research topics of 'Multi-wavelength analytical ultracentrifugation as a tool to characterise protein–DNA interactions in solution'. Together they form a unique fingerprint.

Cite this