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Protein Science (2001), 10:12-16.
Copyright © 2001 The Protein Society

Second virial coefficients as a measure of protein–osmolyte interactions

Gresham T. Weatherly and Gary J. Pielak

Department of Chemistry, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599, USA

Reprint requests to: Gary J. Pielak, Department of Chemistry, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599, USA; e-mail: gary_pielak{at}unc.edu; fax 919-966-3675.

The cytoplasm contains high concentrations of cosolutes. These cosolutes include macromolecules and small organic molecules called osmolytes. However, most biophysical studies of proteins are conducted in dilute solutions. Two broad classes of models have been used to describe the interaction between osmolytes and proteins. One class focuses on excluded volume effects, while the other focuses on binding between the protein and the osmolyte. To better understand protein–smolyte interactions, we have conducted sedimentation equilibrium analytical ultracentrifugation experiments using ferricytochrome c as a model protein. From these experiments, we determined the second virial coefficients for a series of osmolytes. We have interpreted the second virial coefficient as a measure of both excluded volume and protein–osmolyte binding. We conclude that simple models are not sufficient to understand the interactions between osmolytes and proteins.

Keywords: Excluded volume; second virial coefficient; cytochrome c; osmolytes; analytical ultracentrifugation


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