Interactions of insolubilized lectins with membrane glycoproteins in presence of detergents.
The effects of several detergents commonly used to solubilize membrane glycoproteins have been investigated on the binding of hepatoma cell surface [3H]-galactoglycoproteins to, and their elution from, concanavalin A or Ricinus communis lectins conjugated to Sepharose 4B. The optimum conditions (pH, ionic strength) in the presence of ionic [sodium deoxycholate (DOC) and sodium dodecyl sulphate (SDS)] and non-ionic detergents (Triton X-100) at a constant concentration were determined in order to ascertain which would yield the better efficiency. The effects of different detergent concentrations on binding and elution were then studied. The range of concentrations for each detergent to be used without modifying efficiency was determined. Triton X-100 and DOC (0.1-1%) did not change the efficiency on Ricinus lectin-Sepharose, whereas SDS, at a concentration greater than 0.05%, caused a dramatic decrease in efficiency. On concanavalin A-Sepharose, by contrast, the non-ionic detergent had no effect on the efficiency at all the concentrations tested (0.1-1%), while concentrations of more than 0.5% DOC and 0.1% SDS significantly decreased both binding and elution.
D000818:Animals; D006528:Carcinoma, Hepatocellular / Q000378:metabolism; D002846:Chromatography, Affinity / Q000379:methods*; D003208:Concanavalin A / Q000378:metabolism; D003902:Detergents; D006023:Glycoproteins / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D037102:Lectins / Q000378:metabolism; D008113:Liver Neoplasms / Q000378:metabolism; D008565:Membrane Proteins / Q000032:analysis / Q000378:metabolism*; D011485:Protein Binding
D003902:Detergents; D006023:Glycoproteins; D037102:Lectins; D008565:Membrane Proteins; D003208:Concanavalin A