Journal article

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

Christopher R Horne, Amy Henrickson, Borries Demeler, Renwick CJ Dobson

EUROPEAN BIOPHYSICS JOURNAL WITH BIOPHYSICS LETTERS | SPRINGER | Published : 2020

Abstract

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 multim..

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Grants

Awarded by New Zealand Royal Society Marsden Fund


Awarded by Ministry of Business, Innovation and Employment Smart Ideas Grant


Awarded by National Institutes of Health


Awarded by Canada Foundation for Innovation


Awarded by NIH


Awarded by NSF/XSEDE


Awarded by University of Texas


Funding Acknowledgements

We are grateful to the New Zealand Royal Society Marsden Fund (RCJD, UOC1506); Ministry of Business, Innovation and Employment Smart Ideas Grant (RCJD, UOCX1706); and the National Institutes of Health Grants (BD, GM120600 and NSF-ACI-1339649). We acknowledge grant support for CRH (Maurice Wilkins Centre). MWL-AUC experiments were performed at the Canadian Center for Hydrodynamics, University of Lethbridge with support from the Canada Foundation for Innovation Grant CFI-37589. UltraScan multi-wavelength development is supported by NIH Grant GM120600 (BD), UltraScan supercomputer calculations were supported through NSF/XSEDE Grant TG-MCB070039N (BD), and University of Texas Grant TG457201 (BD).