Direct analysis of sedimentation equilibrium distributions reflecting macromolecular interactions

  • P. R. Wills
  • M. P. Jacobsen
  • D. J. WinzorEmail author
Keynote Lectures
Part of the Progress in Colloid & Polymer Science book series (PROGCOLLOID, volume 107)


This review of the characterization of protein interactions by sedimentation equilibrium emphasizes procedures that entail direct determination of the thermodynamic activity of the smallest species contributing to the concentration distribution for that constituent. This approach, which has been regarded as an Australasian oddity for over two decades, is first illustrated by evaluating the association constant for α-chymotrypsin dimerization by the original omega analysis and subsequent refinements thereof. Notable in that regard is the introduction of the psi function, which has evolved from its omega counterpart. Application of the corresponding approach to sedimentation distributions for mixtures of ovalbumin and cytochrome c is presented to illustrate the potential of the psi function for characterizing interactions between dissimilar macromolecular reactants. Also discussed is the means by which these direct analyses of sedimentation equilibrium distributions afford realistic allowance for effects of thermodynamic nonideality on the statistical-mechanical basis of excluded volume.

Key words

Protein self-association heterogeneous association sedimentation equilibrium thermodynamic nonideality 


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  1. 1.
    Adams ET, Williams JW (1964) J Am Chem Soc 86:3454–3461CrossRefGoogle Scholar
  2. 2.
    Roark DE, Yphantis DA (1969) Ann NY Acad Sci 164:245–278PubMedCrossRefGoogle Scholar
  3. 3.
    Hoagland VD, Teller DC (1969) Biochemistry 8:594–602PubMedCrossRefGoogle Scholar
  4. 4.
    Bucci E (1986) Biophys Chem 24:47–52PubMedCrossRefGoogle Scholar
  5. 5.
    Chatelier RC, Minton AP (1987) Biopolymers 26:507–524PubMedCrossRefGoogle Scholar
  6. 6.
    Minton AP (1990) Anal Biochem 190:1–6PubMedCrossRefGoogle Scholar
  7. 7.
    Milthorpe BK, Jeffrey PD, Nichol LW (1975) Biophys Chem 3:169–176PubMedCrossRefGoogle Scholar
  8. 8.
    Johnson ML, Correia JJ, Yphantis DA, Halvorson HR (1981) Biophys J 36:575–588PubMedGoogle Scholar
  9. 9.
    Wills PR, Jacobsen MP, Winzor DJ (1996) Biopolymers 38:119–130CrossRefGoogle Scholar
  10. 10.
    Tellam R, de Jersey J, Winzor DJ (1979) Biochemistry 24:5316–5321CrossRefGoogle Scholar
  11. 11.
    Tellam R, Winzor DJ (1980) Arch Biochem Biophys 201:20–34PubMedCrossRefGoogle Scholar
  12. 12.
    Johnson ML, Straume M (1994) In: Schuster TM, Laue TM (eds) Modern Analytical Ultracentrifugation. Birkhäuser, Boston, pp 37–65Google Scholar
  13. 13.
    Williams JW, Van Holde KE, Baldwin RL, Fujita H (1958) Chem Revs 58:715–806CrossRefGoogle Scholar
  14. 14.
    Wills PR, Winzor DJ (1992) In: Harding SE, Rowe AJ, Horton JC (eds) Analytical Ultracentrifugation in Biochemistry and Polymer Science. Roy Soc Chem, Cambridge, pp 311–330Google Scholar
  15. 15.
    Wills PR, Comper WD, Winzor DJ (1993) Arch Biochem Biophys 300:206–212PubMedCrossRefGoogle Scholar
  16. 16.
    Ogston AG, Winzor DJ (1975) J Phys Chem 79:2496–2500CrossRefGoogle Scholar
  17. 17.
    Yphantis DA (1964) Biochemistry 3:297–317PubMedCrossRefGoogle Scholar
  18. 18.
    Morris M, Ralston GB (1985) Biophys Chem 23:49–61PubMedCrossRefGoogle Scholar
  19. 19.
    Jacobsen MP, Winzor DJ (1992) Biophys Chem 45:119–132CrossRefGoogle Scholar
  20. 20.
    Wills PR, Jacobsen MP, Winzor DJ (1996) Biopolymers 38:119–130CrossRefGoogle Scholar
  21. 21.
    Adams ET, Fujita H (1963) In: Williams JW (ed) Ultracentrifugal Analysis in Theory and Experiment. Academic Press, New York, pp 119–128Google Scholar
  22. 22.
    Wills PR, Siezen RJ, Nichol LW (1980) Biophys Chem 11:71–82PubMedCrossRefGoogle Scholar
  23. 23.
    Jeffrey PD (1981) In: Frieden C, Nichol LW (eds) Protein-Protein Interactions. Wiley, New York, pp 213–256Google Scholar
  24. 24.
    Harris SJ, Winzor DJ (1988) Arch Biochem Biophys 265:458–465PubMedCrossRefGoogle Scholar
  25. 25.
    Winzor DJ, Wills PR (1994) In: Schuster TM, Laue TM (eds) Modern Analytical Ultracentrifugation. Birkhäuser, Boston, pp 66–80Google Scholar
  26. 26.
    Hill TL, Chen YD (1973) Biopolymers 12:1285–1312CrossRefGoogle Scholar
  27. 27.
    Aune KC, Goldsmith LC, Timasheff SN (1971) Biochemistry 10:1617–1621PubMedCrossRefGoogle Scholar
  28. 28.
    Shearwin KE, Winzor DJ (1988) Biophys Chem 31:287–294PubMedCrossRefGoogle Scholar
  29. 29.
    Wills PR, Winzor DJ (1993) Biopolymers 33:1627–1629CrossRefGoogle Scholar
  30. 30.
    Winzor DJ, Wills PR (1995) In: Gregory RB (ed) Protein-Solvent Interactions Dekker, New York, pp 483–520Google Scholar
  31. 31.
    Cohen G, Eisenberg H (1968) Biopolymers 6:1077–1100PubMedCrossRefGoogle Scholar
  32. 32.
    Reisler E, Eisenberg H (1969) Biochemistry 8:4572–4578PubMedCrossRefGoogle Scholar
  33. 33.
    Lee JC, Timasheff SN (1981) J Biol Chem 256:7193–7201PubMedGoogle Scholar
  34. 34.
    Timasheff SN (1995) In: Gregory RB (ed) Protein-Solvent Interactions, Dekker, New York, pp 445–482Google Scholar
  35. 35.
    Jacobsen MP, Wills PR, Winzor DJ (1996) Biochemistry 35:13 173–13 179Google Scholar
  36. 36.
    Casassa EF, Eisenberg H (1964) Adv Protein Chem 19:287–395PubMedCrossRefGoogle Scholar
  37. 37.
    Eisenberg H (1976) Biological Macromolecules and Polyelectrolytes in Solution. Clarendon Press, Oxford.Google Scholar
  38. 38.
    Derrien Y, Michel R, Pedersen KO, Roche J (1949) Biochim Biophys Acta 3:436–441CrossRefGoogle Scholar
  39. 39.
    Steiner RF, Edelhoch H (1961) J Am Chem Soc 83:1435–1444CrossRefGoogle Scholar
  40. 40.
    Lee JC, Timasheff SN (1974) Arch Biochem Biophys 165:268–273PubMedCrossRefGoogle Scholar
  41. 41.
    Prakash V, Timasheff SN (1981) Anal Biochem 117:330–335PubMedCrossRefGoogle Scholar
  42. 42.
    Taylor JF, Lowry C (1956) Biochim Biophys Acta 20:109–117PubMedCrossRefGoogle Scholar
  43. 43.
    Nichol LW, Ogston AG (1965) J Phys Chem 69:4365–4367CrossRefGoogle Scholar
  44. 44.
    Nichol LW, Jeffrey PD, Milthorpe BK (1976) Biophys Chem 4:259–267PubMedCrossRefGoogle Scholar
  45. 45.
    Jeffrey PD, Nichol LW, Teasdale RD (1979) Biophys Chem 10:379–387PubMedCrossRefGoogle Scholar
  46. 46.
    Cölfen H, Harding SE, Vårum KM, Winzor DJ (1996) Carbohydrate Polym 30:45–53CrossRefGoogle Scholar
  47. 47.
    Laue TM, Senear DF, Eaton S, Ross AJB (1993) Biochemistry 32:2469–2472PubMedCrossRefGoogle Scholar
  48. 48.
    Kim T, Tsuykiyama T, Lewis MS, Wu C (1994) Protein Sci 3:1040–1051PubMedGoogle Scholar
  49. 49.
    Bailey MF, Davidson BE, Minton AP, Sawyer WH, Howlett GJ (1996) J Mol Biol 263:671–684PubMedCrossRefGoogle Scholar
  50. 50.
    Laue TM, Hazard AL, Ridgeway TM, Yphantis DA (1989) Anal Biochem 182:377–382PubMedCrossRefGoogle Scholar
  51. 51.
    Creeth JM, Winzor DJ (1962) Biochem J 83:566–574PubMedGoogle Scholar

Copyright information

© Dr. Dietrich Steinkopff Verlag GmbH & Co. KG 1997

Authors and Affiliations

  1. 1.Department of PhysicsUniversity of AucklandAucklandNew Zealand
  2. 2.Centre for Protein Structure, Function and Engineering Department of BiochemistryUniversity of QueenslandBrisbaneAustralia

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