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Eur Biophys J. 2018 Apr;47(3):291-295. doi: 10.1007/s00249-017-1259-x. Epub 2017 Oct 4.

Allowance for radial dilution in evaluating the concentration dependence of sedimentation coefficients for globular proteins.

Author information

1
Department of Chemistry and Biochemistry, Alberta RNA Research and Training Institute, University of Lethbridge, 4401 University Drive, Lethbridge, AB, T1K 3M4, Canada. trushar.patel@uleth.ca.
2
Discovery Lab, Medical Sciences Building, University of Alberta, Edmonton, AB, T6G 2H7, Canada. trushar.patel@uleth.ca.
3
Department of Microbiology, Immunology and Infectious Diseases, Cumming School of Medicine, University of Calgary, Calgary, AB, T2N 4N1, Canada. trushar.patel@uleth.ca.
4
School of Chemistry and Molecular Biosciences, University of Queensland, Brisbane, QLD, 4072, Australia.
5
National Center for Macromolecular Hydrodynamics, School of Biosciences, University of Nottingham, Sutton Bonington Campus, Loughborough, LE2 5RD, UK. david.scott@nottingham.ac.uk.
6
ISIS Spallation Neutron and Muon Source, Rutherford Appleton Laboratory, Harwell Innovation Campus, Oxfordshire, OX11 OFA, UK. david.scott@nottingham.ac.uk.
7
Research Complex at Harwell, Rutherford Appleton Laboratory, Harwell Innovation Campus, Oxfordshire, OX11 OFA, UK. david.scott@nottingham.ac.uk.

Abstract

The accuracy with which the concentration dependence of the sedimentation coefficient, s = s 0(1 - kc), can be quantified for globular proteins by commonly used procedures has been examined by subjecting simulated sedimentation velocity distributions for ovalbumin to c(s)‒s analysis. Because this procedure, as well as its g(s)‒s counterpart, is based on assumed constancy of s over the time course of sedimentation coefficient measurement in a given experiment, the best definition of the concentration coefficient k is obtained by associating the measured s with the mean of plateau concentrations for the initial and final distributions used for its determination. The return of a slightly underestimated k (by about 3%) is traced to minor mislocation of the air‒liquid meniscus position as the result of assuming time independence of s in a given experiment. Although more accurate quantification should result from later SEDFIT and SEDANAL programs incorporating the simultaneous evaluation of s 0 and k, the procedures based on assumed constancy of s suffice for determining the limiting sedimentation coefficient s 0-the objective of most s‒c dependence studies.

KEYWORDS:

Concentration dependence; Sedimentation coefficient; Sedimentation velocity; Ultracentrifugation

PMID:
28980105
DOI:
10.1007/s00249-017-1259-x
[Indexed for MEDLINE]

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