Ultraviolet difference spectroscopic analysis of the saccharide-binding properties of Ricinus communis agglutinin.
The nature of the binding of saccharides to Ricinus communis agglutinin was studied by ultraviolet difference spectroscopy. Upon binding of galactose and galactose-containing saccharides, R. communis agglutinin displayed difference spectra with an extreme maximum at 291-293 nm and a smaller maximum at 284-285 nm. Such difference spectra suggest that the environment of a tryptophan residue located at or near the saccharide-binding site of R. communis agglutinin is being changed by an interaction between a tryptophan residue and the bound saccharides. The value of the difference spectra (delta epsilon) increased upon progressive addition of saccharide until the saccharide binding site was saturated with ligand. From the increase in delta epsilon at 291-293 nm, the association constants were obtained for the R. communis agglutinin-saccharide interaction over the temperature range 5-35 degrees C and various pH values. The results clearly demonstrate that the association constants are nearly equal in the range of pH 5-8, but decrease beyond the above pH range and with elevation of temperature. From the thermodynamic parameters for the binding of various saccharides to R. communis agglutinin, we suggest that there exists a subsite structure in the saccharide-binding site of the R. communis agglutinin molecule.
D001665:Binding Sites; D050260:Carbohydrate Metabolism; D005690:Galactose / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007785:Lactose / Q000378:metabolism; D037102:Lectins; D008756:Methylgalactosides / Q000378:metabolism; D037121:Plant Lectins; D011887:Raffinose / Q000378:metabolism; D013056:Spectrophotometry, Ultraviolet; D013696:Temperature; D014364:Tryptophan
D037102:Lectins; D008756:Methylgalactosides; D037121:Plant Lectins; C017707:Ricinus communis agglutinin-1; D014364:Tryptophan; D007785:Lactose; D011887:Raffinose; D005690:Galactose