Neoglycoconjugates of mannan with bovine serum albumin and their interaction with lectin concanavalin A.
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: (i) 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 x L(-1) (close to K(D) = 10(-8) mol x 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.
Mislovicová
D
D
Institute of Chemistry, Slovak Academy of Sciences, SK-842 38 Bratislava, Slovakia. chemmisl@savba.sk
Masárová
J
J
Svitel
J
J
Mendichi
R
R
Soltés
L
L
Gemeiner
P
P
Danielsson
B
B
D003208:Concanavalin A / Q000737:chemistry*; D006001:Glycoconjugates / Q000138:chemical synthesis* / Q000494:pharmacology*; D008351:Mannans / Q000737:chemistry* / Q000494:pharmacology*; D008970:Molecular Weight; D010084:Oxidation-Reduction; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000378:metabolism; D012710:Serum Albumin, Bovine / Q000737:chemistry*; D020349:Surface Plasmon Resonance
D006001:Glycoconjugates; D008351:Mannans; D003208:Concanavalin A; D012710:Serum Albumin, Bovine