Probing protein aggregation by time-resolved fluorescence during beta-lactoglobulin crystal growth

Eur Biophys J. 2002 May;31(2):111-7. doi: 10.1007/s00249-002-0208-4. Epub 2002 Apr 11.

Abstract

We have used the fluorescence anisotropy (FA) decay of retinol bound to bovine beta-lactoglobulin to monitor the time evolution of protein aggregation during the early stages of crystal growth. With this approach we have followed the formation of aggregates at different concentrations of ammonium sulfate, the precipitant used for crystallization. The average aggregation number is found to depend on precipitant concentration, and to be restricted to small numbers ranging from 2 to 5, also in the presence of visible growing crystals. The effect of particle distribution and of low probe-to-protein saturation on the FA response is also discussed in detail.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Ammonium Sulfate / pharmacology
  • Animals
  • Anisotropy
  • Cattle
  • Crystallization
  • Dimerization
  • Lactoglobulins / chemistry*
  • Light
  • Models, Statistical
  • Scattering, Radiation
  • Spectrometry, Fluorescence / methods*
  • Time Factors
  • Vitamin A / chemistry

Substances

  • Lactoglobulins
  • Vitamin A
  • Ammonium Sulfate