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Carbohydrate-Protein Interactions at Single 20 nm-Beads:

In recent years, the emerging field of glycobiology or glycomics has attracted a considerable amount of attention because of its vital importance for the understanding of many diseases. Carbohydrates such as glycoproteins, glycolipids, glycopeptides, glycosaminoglycans or other glycoconjugates have long been known to take part in a multitude of biological processes. Examples are immune response, inflammation and viral infection, cell–cell interactions, signal transduction and developmental biology. However, since oligosaccharides are structurally even more complex than proteins or nucleic acids there is still a great need for suitable biophysical methods to elucidate specific carbohydrate-biomolecule interactions. In this project we investigate carbohydrate-protein interactions by exploration of the aggregation behaviour of carbohydrated fluorescing nanospheres of 20 nm diameter in the presence of carbohydrate-binding proteins via two-photon fluorescence correlation spectroscopy on a single particle level. The assay principle allows a determination of relative protein-carbohydrate binding affinities without labelling the proteins and may serve as a simple model system for the physical and chemical interactions of proteins with carbohydrates, for example, at cell or virus surfaces.

Schematical view of the polymerization processes during the aggregation of the carbohydrated 20 nm spheres (circles with grey pins) in the presence of different concentrations of carbohydrate-binding proteins (red symbols).

For details see:

W. Pohl, H. Hellmuth, M. Hilbert, J. Seibel and Peter J. Walla, "A two-photon fluorescence correlation study of lectins interacting with carbohydrated 20 nm beads”, ChemBioChem 7, 268-274, (2006).

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