Biotinylated diaminopyridine: an approach to tagging oligosaccharides and exploring their biology.

AUTOR(ES)

Rothenberg, B E

RESUMO

Fluorescent tagging of free oligosaccharides by reductive amination permits sensitive detection and fractionation of these molecules. To expand the scope of this approach, we have synthesized a fluorescent reagent, 2-amino-(6-amidobiotinyl)pyridine. This reagent can tag oligosaccharides under nondegradative conditions with high efficiency. The resulting adducts show excellent fractionation by reverse-phase HPLC with sensitive detection in the low picomole range. When combined with sequential exoglycosidase digestion, stepwise sequencing of the sugar chains is possible. The biotinyl group can also be used to recover the sugar chain from reaction mixtures. The high-affinity interaction of the biotinyl group with multivalent avidin or streptavidin can be used to create the functional equivalent of neoglycoproteins carrying multiple copies of oligosaccharides of defined structure. These complexes allow the production of IgG antibodies directed against the oligosaccharide chain. They can also harness the power of (strept)avidin-biotin technology for the detection and isolation of oligosaccharide-specific receptors from native sources of recombinant libraries.

Documentos Relacionados

• Non-Isotopic Methods in Molecular Biology. A Practical Approach
• “Translational” Legume Biology. Models to Crops
• Viral pathogenesis and molecular biology.
• Perspectives series: cell adhesion in vascular biology. Integrin signaling in vascular biology.
• Biomechanical activation: an emerging paradigm in endothelial adhesion biology.