Glycoprotein Nature of Glycosidases from Leaves ofPisum sativumL

Publisher Website
Google Scholar
Abstract
α-Mannosidase, β-N-acetylglucosaminidase, α- and β-galactosidase and β-glucosidase were partially purified from leaves of Pisum sativum by ammonium sulphate fractionation and column chromatography on DEAE-Sephadex A-50 and hydroxylapatite. At least two molecular forms of each enzyme were resolved by these techniques except for β-glucosidase of which only one form was resolved. Except for one form of α-galactosidase, all of the glycosidases thus purified were completely bound by Sepharose-linked Concanavalin A. The binding was strongly inhibited by cr-methyl-D-mannoside and no binding to Sepharose-6-B occurred indicating that these glycosidases contain mannose-rich oligosaccharides. The glycoprotein nature of α-mannosidase, β-galactosidase and β-glucosidase was further demonstrated by chromatography on phenylboronate agarose columns. The differences in the concentration of cr-methyl-D-mannoside and sorbitol required to elute the various glycosidases from Sepharose-linked Con A and phenylboronate agarose, respectively, suggested that these enzymes are glycosylated to various degrees or that structural variation in their carbohydrate moieties occur. This is the first demonstration that glycosylation of several glycosidases present in a single plant species is apparently a generalized feature of these enzymes.

This publication has 1 reference indexed in Scilit: