Dynamic light scattering: a practical guide and applications in biomedical sciences
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Dynamic light scattering (DLS), also known as photon correlation spectroscopy (PCS), is a very powerful tool for studying the diffusion behaviour of macromolecules in solution. The diffusion coefficient, and hence the hydrodynamic radii calculated from it, depends on the size and shape of macromolecules. In this review, we provide evidence of the usefulness of DLS to study the homogeneity of proteins, nucleic acids, and complexes of protein–protein or protein–nucleic acid preparations, as well as to study protein–small molecule interactions. Further, we provide examples of DLS’s application both as a complementary method to analytical ultracentrifugation studies and as a screening tool to validate solution scattering models using determined hydrodynamic radii.
KeywordsAnalytical ultracentrifuge Diffusion coefficient Dynamic light scattering Hydrodynamic radius Light scattering Protein–ligand interactions Protein–nucleic acid complexes Protein–protein complexes
TRP acknowledges Manitoba Institute of Child Health (2008–10) and Canadian Institutes of Health Research (2010–12) for postdoctoral fellowship awards, and Marie Skłodowska-Curie Fellowship program (2013–15) for their financial support. JS holds a Canada Research Chair in Structure Biology and was supported by funding from NSERC. SAM is supported by NSERC Discovery grant (RGPIN-2015-06142). The authors congratulate Prof. Donald Winzor on his 80th birthday and thank him for his immense contributions in the development and applications of biophysical methods. Dr. Patel thanks Prof. Winzor for being his mentor. The authors thank Drs. Hall and Harding for an opportunity to contribute in a special issue “Analytical Quantitative Relations in Biochemistry”.
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Conflict of Interests
Jörg Stetefeld declares that he has no conflicts of interest. Sean A. McKenna declares that he has no conflicts of interest. Trushar R. Patel declares that he has no conflicts of interest.
This article does not contain any studies with human participants or animals performed by any of the authors.
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