Influence of mannan epitopes in glycoproteins--Concanavalin A interaction. Comparison of natural and synthetic glycosylated proteins.
Two natural glycoproteins/glycoenzymes, invertase and glucoamylase, and two neoglycoconjugates, synthetized from Saccharomyces cerevisiae mannan, bovine serum albumin and penicillin G acylase were tested for interaction with lectin Concanavalin A (Con A). The interaction of natural and synthetic glycoproteins with Con A was studied using three different experimental methods: (i). quantitative precipitation in solution (ii). sorption to Con A immobilized on bead cellulose; and (iii). kinetic measurement of the interaction by surface plasmon resonance. Prepared neoglycoproteins were further characterized: saccharide content, molecular weight, polydispersion, kinetic and equilibrium association constants with Con A were determined. It can be concluded that the used conjugation method proved to be able to produce neoglycoproteins with similar properties like natural glycoproteins, i.e. enzymatic activity (protein part) and lectin binding activity (mannan part) were preserved and the neoglycoconjugates interact with Con A similarly as natural mannan-type glycoproteins.
Mislovicová
D
D
Institute of Chemistry, Slovak Academy of Sciences, SK-842 38, Bratislava, Slovakia. chemmisl@savba.sk
Masárová
J
J
Svitel
J
J
Gemeiner
P
P
D001234:Aspergillus niger / Q000201:enzymology; D001665:Binding Sites; D003208:Concanavalin A / Q000493:pharmacokinetics*; D000939:Epitopes / Q000737:chemistry / Q000378:metabolism; D005087:Glucan 1,4-alpha-Glucosidase / Q000378:metabolism; D006001:Glycoconjugates / Q000737:chemistry / Q000378:metabolism; D006023:Glycoproteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D006026:Glycoside Hydrolases / Q000378:metabolism; D006031:Glycosylation; D008351:Mannans / Q000737:chemistry* / Q000378:metabolism; D010399:Penicillin Amidase / Q000378:metabolism; D012441:Saccharomyces cerevisiae; D043324:beta-Fructofuranosidase
D000939:Epitopes; D006001:Glycoconjugates; D006023:Glycoproteins; D008351:Mannans; D003208:Concanavalin A; D006026:Glycoside Hydrolases; D043324:beta-Fructofuranosidase; D005087:Glucan 1,4-alpha-Glucosidase; D010399:Penicillin Amidase