Metabolic Inhibition of Sialyl-Lewis X Biosynthesis by 5-Thiofucose Remodels the Cell Surface and Impairs Selectin-Mediated Cell Adhesion*
| Autor: | Wesley F. Zandberg, David J. Vocadlo, Jayakanthan Kumarasamy, B. Mario Pinto |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2012 |
| Předmět: |
Fucosyltransferase
Glycosylation Cell Glycobiology and Extracellular Matrices Oligosaccharides CHO Cells Biology Biochemistry chemistry.chemical_compound Cricetulus Cricetinae medicine Cell Adhesion Animals Humans Enzyme Inhibitors Cell adhesion Sialyl Lewis X Antigen Molecular Biology Fucose Cell adhesion molecule Endothelial Cells Cell Biology Hep G2 Cells Fucosyltransferases Extravasation Cell biology carbohydrates (lipids) medicine.anatomical_structure Sialyl-Lewis X chemistry Metabolic Engineering Cancer cell biology.protein Selectins Selectin |
| Popis: | Sialyl-Lewis X (sLe(X)) is a tetrasaccharide that serves as a ligand for the set of cell adhesion proteins known as selectins. This interaction enables adhesion of leukocytes and cancer cells to endothelial cells within capillaries, resulting in their extravasation into tissues. The last step in sLe(X) biosynthesis is the α1,3-fucosyltrasferase (FUT)-catalyzed transfer of an L-fucose residue to carbohydrate acceptors. Impairing FUT activity compromises leukocyte homing to sites of inflammation and renders cancer cells less malignant. Inhibition of FUTs is, consequently, of great interest, but efforts to generate glycosyltransferase inhibitors, including FUT inhibitors, has proven challenging. Here we describe a metabolic engineering strategy to inhibit the biosynthesis of sLe(X) in cancer cells using peracetylated 5-thio-L-fucose (5T-Fuc). We show that 5T-Fuc is taken up by cancer cells and then converted into a sugar nucleotide analog, GDP-5T-Fuc, that blocks FUT activity and limits sLe(X) presentation on HepG2 cells with an EC(50) in the low micromolar range. GDP-5T-Fuc itself does not get transferred by either FUT3 or FUT7 at a measurable rate. We further demonstrate that treatment of cells with 5T-Fuc impaired their adhesive properties to immobilized adhesion molecules and human endothelial cells. 5T-Fuc, therefore, is a useful probe that can be used to modulate sLe(X) levels in cells to evaluate the consequences of inhibiting FUT-mediated sLe(X) formation. These data also reveal the utility of using sugar analogues that lead to formation of donor substrate analogues within cells as a general approach to blocking glycosyltransferases in cells. |
| Databáze: | OpenAIRE |
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