Evidence for the glycoprotein nature of radish beta-fructosidase.
Concanavalin A (Con A) was utilized free, bound to Sepharose 4 B or cross-linked to glutaraldehyde to investigate the possibility of binding this lectin to radish beta-fructosidase (E.C.3.2.1.26). The choice of cross-linked Con A as affinoadsorbent is discussed and standard conditions for binding are defined. Specificity of precipitation of this enzyme by the lectin was especially investigated. Thus, the possibility of binding was tested in the presence of high ionic strength, ethylene glycol, alpha-methyl mannoside, alpha-methyl glucoside and during periodate oxidation of the enzyme. Based on the interactions observed between beta-fructosidase and Con A under these conditions it is concluded that the saccharide binding site of the lectin is primarily involved with a secondary contribution from the hydrophobic site. The specificity of binding and the complete precipitation of beta-fructosidase activity by the insolubilized lectin imply that all beta-fructosidase activity measured in Raphanus sativus seedling extracts is linked to (a) glycoprotein form(s) of this enzyme.
D003208:Concanavalin A; D004800:Enzymes, Immobilized / Q000378:metabolism; D005632:Fructose; D006023:Glycoproteins; D006026:Glycoside Hydrolases / Q000378:metabolism*; D007700:Kinetics; D037121:Plant Lectins; D010944:Plants / Q000201:enzymology; D011485:Protein Binding
D004800:Enzymes, Immobilized; D006023:Glycoproteins; D037121:Plant Lectins; D003208:Concanavalin A; D005632:Fructose; D006026:Glycoside Hydrolases