Identification of Endothelial Cell Surface Carbohydrate-Binding Receptors by Carbohydrate Ligand Mimicry Peptides
Substantial evidence suggests that cell surface carbohydrate antigens, particularly those containing fucose residues, are related to cancer malignancy. To investigate the mechanisms underlying cell surface carbohydrate-dependent cancer metastasis, we developed a 7-mer peptide, designated as I-peptide, which is capable of inhibiting FUT3-transfected B16 cell colonization of lung in the mouse. I-peptide inhibited lung colonization of FUT3-B16 cells in E-/P-selectin doubly deficient mutant mice, suggesting the existence of an as yet unidentified carbohydrate-binding endothelial receptor distinct from E-/P-selectins. Subsequently, we isolated the endothelial receptors by I-peptide affinity chromatography and identified them by proteomics. Thus, I-peptide receptors were identified to be pre-mRNA splicing factor (Sfrs) proteins. We also found a fragment of annexin A1 (Anxa1) bound to I-peptide. Our study demonstrated the usefulness of carbohydrate-ligand mimicry peptide for the identification of novel carbohydrate-binding proteins.
KeywordsCarbohydrate EDTA Carboxylate Electrophoresis Luminal
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