Protein hydration varies with protein crowding and with applied pressure: a sedimentation velocity study

  • N. Errington
  • P. Mistry
  • A. J. Rowe
Conference paper
Part of the Progress in Colloid and Polymer Science book series (PROGCOLLOID, volume 119)


We have recently shown, using Derjaguin—Landau—Verwey—Overbeek theory, that the stability of protein solutions can be accounted for primarily in terms of the energy barrier presented by the bound water surrounding the protein, rather than by net repulsive forces. In further work we demonstrated experimentally, using precision densimetry and dynamic light scattering, that the amount of this water bound to proteins varies with temperature. We have now used the analytical ultracentrifuge (AUC) to study possible effects of crowding and applied hydrostatic pressure on water binding and hence on the sedimentation velocity of proteins. We show that whilst self-cancelling of effects minimises changes in the s values, there are predictions, which we are able to confirm experimentally, that both high protein concentration and elevated hydrostatic pressure at levels found in the AUC will lead to effects attributable to additional hydration. It is concluded that protein hydration is, over a range of conditions, a variable rather than a constant quantity. This finding is significant in relation to the stability and formulation of protein solutions.

Key words

Hydration Sedimentation velocity Pressure Crowding Stability of protein solutions 


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  1. 1.
    Rowe AJ (2001) Biophys Chem (in press)Google Scholar
  2. 2.
    Schuck P (1998) Biophys J 75:1503–1512Google Scholar
  3. 3.
    Rowe AJ (1977) Biopolymers 16:2595–2611CrossRefGoogle Scholar
  4. 4.
    Rowe AJ (1992) In: Harding SE, Rowe AJ, Horton JC (eds) Analytical ultra-centrifugation in biochemistry, poly mer science. Royal Society of Chemistry, Cambridge, pp 394–406Google Scholar
  5. 5.
  6. 6.
    Gross M, Jaenicke R (1994) Eur J Biochem 221:617–630CrossRefGoogle Scholar
  7. 7.
    Cho CH, Urquidi J, Robinson GW (1999) J Chem Phys 111:10171–10174CrossRefGoogle Scholar
  8. 8.
    Philo JS (1977) Biophys J 72:435–444CrossRefGoogle Scholar
  9. 9.
    Philo JS (2000) Anal Biochem 279:151–163CrossRefGoogle Scholar
  10. 10.
    van Holde KE, Weischet WO (1978) Biopolymers 17:1387–1403CrossRefGoogle Scholar
  11. 11.
    Timasheff SN (1999) Adv Protein Chem 51:355–432CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2002

Authors and Affiliations

  • N. Errington
    • 1
  • P. Mistry
    • 2
  • A. J. Rowe
    • 1
  1. 1.NCMH Business Centre School of BiosciencesUniversity of NottinghamLeicestershireUK
  2. 2.Department of BiochemistryUniversity of LeicesterLeicesterUK

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