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- Mislovicova, D, et al.
(författare)
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Influence of mannan epitopes in glycoproteins - Concanavalin A interaction. Comparison of natural and synthetic glycosylated proteins
- 2002
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Ingår i: International Journal of Biological Macromolecules. - 1879-0003. ; 30:5, s. 251-258
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Tidskriftsartikel (refereegranskat)abstract
- 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.
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2. |
- Mislovicova, D, et al.
(författare)
-
Neoglycoconjugates of mannan with bovine serum albumin and their interaction with lectin concanavalin A
- 2002
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Ingår i: Bioconjugate Chemistry. - : American Chemical Society (ACS). - 1520-4812 .- 1043-1802. ; 13:1, s. 136-142
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Tidskriftsartikel (refereegranskat)abstract
- Neoglycoconjugates were prepared from mannan isolated from yeast Saccharomyces cerevisiae and activated by periodate oxidation to create aldehyde groups. Various degrees of oxidation introduced 11-28 aldehyde groups per mannan molecule and simultaneously resulted in a molar mass decrease from 46 to 44.5-31 kDa. The activated mannans were subsequently conjugated with bovine serum albumin forming neoglycoconjugates. Some parameters of these mannan-bovine serum albumin conjugates were characterized: saccharide content 25-30% w/w, molar mass within the range 169-246 kDa, and polydispersion (M-w/M-n) from 2.8 to 3.6. The interaction of these conjugates with lectin concanavalin A was studied using three different methods: W quantitative precipitation in solution; (ii) sorption to concanavalin A immobilized on bead cellulose; and (iii) kinetic measurement of the interaction by surface plasmon resonance. Quantitative precipitation assay showed only negligible differences in the precipitation course of original mannan and the corresponding mannan-bovine serum albumin conjugates. Both the sorption method (equilibrium method) and the surface plasmon resonance measurement (kinetic method) demonstrates that the values of dissociation constant K-D of all synthetic neoglycoconjugates were within the range 10(-7)-10(-8) mol.L-1 (close to K-D = 10(-1) mol-L-1 determined by the sorption method for the original mannan). In conclusion, characterization of synthetic neoglycoconjugates confirmed that the method used for their preparation retained the ability of mannan moiety to interact with concanavalin A.
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