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. 2000 Oct;79(4):2178–2187. doi: 10.1016/S0006-3495(00)76466-6

Analysis of sedimentation equilibrium distributions reflecting nonideal macromolecular associations.

P R Wills 1, M P Jacobsen 1, D J Winzor 1
PMCID: PMC1301108  PMID: 11023922

Abstract

A rigorous statistical-mechanical approach is adopted to derive general quantitative expressions that allow for the effects of thermodynamic nonideality in equilibrium measurements reflecting interaction between dissimilar macromolecular reactants. An analytical procedure based on these expressions is then formulated for obtaining global estimates of equilibrium constants and the corresponding reference thermodynamic activities of the free reactants in each of several sedimentation equilibrium experiments. The method is demonstrated by application to results from an ultracentrifugal study of an electrostatic interaction between ovalbumin and cytochrome c (Winzor, D. J., M. P. Jacobsen, and P. R. Wills. 1998. Biochemistry. 37:2226-2233). It is demonstrated that reliable estimates of relevant thermodynamic parameters are extracted from the data through statistical analysis by means of a simple nonlinear fitting procedure.

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Selected References

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  1. Bailey M. F., Davidson B. E., Minton A. P., Sawyer W. H., Howlett G. J. The effect of self-association on the interaction of the Escherichia coli regulatory protein TyrR with DNA. J Mol Biol. 1996 Nov 15;263(5):671–684. doi: 10.1006/jmbi.1996.0607. [DOI] [PubMed] [Google Scholar]
  2. Haschemeyer R. H., Bowers W. F. Exponential analysis of concentration or concentration difference data for discrete molecular weight distributions in sedimentation equilibrium. Biochemistry. 1970 Jan 20;9(2):435–445. doi: 10.1021/bi00804a035. [DOI] [PubMed] [Google Scholar]
  3. Jacobsen M. P., Wills P. R., Winzor D. J. Thermodynamic analysis of the effects of small inert cosolutes in the ultracentrifugation of noninteracting proteins. Biochemistry. 1996 Oct 8;35(40):13173–13179. doi: 10.1021/bi960939q. [DOI] [PubMed] [Google Scholar]
  4. Johnson M. L., Correia J. J., Yphantis D. A., Halvorson H. R. Analysis of data from the analytical ultracentrifuge by nonlinear least-squares techniques. Biophys J. 1981 Dec;36(3):575–588. doi: 10.1016/S0006-3495(81)84753-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Kim S. J., Tsukiyama T., Lewis M. S., Wu C. Interaction of the DNA-binding domain of Drosophila heat shock factor with its cognate DNA site: a thermodynamic analysis using analytical ultracentrifugation. Protein Sci. 1994 Jul;3(7):1040–1051. doi: 10.1002/pro.5560030706. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Laue T. M., Senear D. F., Eaton S., Ross J. B. 5-hydroxytryptophan as a new intrinsic probe for investigating protein-DNA interactions by analytical ultracentrifugation. Study of the effect of DNA on self-assembly of the bacteriophage lambda cI repressor. Biochemistry. 1993 Mar 16;32(10):2469–2472. doi: 10.1021/bi00061a003. [DOI] [PubMed] [Google Scholar]
  7. Milthorpe B. K., Jeffrey P. D., Nichol L. W. The direct analysis of sedimentation equilibrium results obtained with polymerizing systems. Biophys Chem. 1975 Apr;3(2):169–176. doi: 10.1016/0301-4622(75)80007-x. [DOI] [PubMed] [Google Scholar]
  8. Wills P. R., Hall D. R., Winzor D. J. Interpretation of thermodynamic non-ideality in sedimentation equilibrium experiments on proteins. Biophys Chem. 2000 May 15;84(3):217–225. doi: 10.1016/s0301-4622(00)00124-1. [DOI] [PubMed] [Google Scholar]
  9. Wills P. R., Nichol L. W., Siezen R. J. The indefinite self-association of lysozyme: consideration of composition-dependent activity coefficients. Biophys Chem. 1980 Feb;11(1):71–82. doi: 10.1016/0301-4622(80)85009-5. [DOI] [PubMed] [Google Scholar]
  10. Winzor D. J., Jacobsen M. P., Wills P. R. Direct analysis of sedimentation equilibrium distributions reflecting complex formation between dissimilar reactants. Biochemistry. 1998 Feb 24;37(8):2226–2233. doi: 10.1021/bi972211v. [DOI] [PubMed] [Google Scholar]

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