Binding of galactose and lactose to ricin. Equilibrium studies

Biochim Biophys Acta. 1978 Sep 26;536(1):18-26. doi: 10.1016/0005-2795(78)90047-8.

Abstract

The interaction of ricin, one of the two lectins of Ricinus sanguineus, with its specific ligands galactose and lactose (4-O-beta-D-galactopyranosyl-D-glucopyranose) has been studied by means of equilibrium dialysis, analytical ultracentrifugation and fluorescence polarization. In the studied concentration range, only one molecule of galactose is bound per molecule of ricin with an association constant, Ka = 6900 m-1 at 4 degrees C. Scatchard plots of equilibrium dialysis data show that two molecules of lactose bind to one molecule of ricin, without modification of molecular weight of the lectin. Together with results of microcalorimetric experiments and agglutination of erythrocytes by ricin, equilibrium dialysis data indicate that the lectin contains two distinct saccharide binding sites. Regardless of the existence of extended sites, it is not possible to select between the two models: (a) two independent sites (Ka1 = 35 000 M-1, Ka2 = 2800 M-1 at 4 degrees C) or (b) two identical sites with negative cooperativity.

MeSH terms

  • Dialysis
  • Galactose*
  • Kinetics
  • Lactose*
  • Protein Binding
  • Ricin*
  • Spectrometry, Fluorescence
  • Thermodynamics

Substances

  • Ricin
  • Lactose
  • Galactose