The heparan sulfate motif (GlcNS6S-IdoA2S)3, common in heparin, has a strict topography and is involved in cell behavior and disease.
The Journal of biological chemistry
Amyloidogenic Proteins; Amyloidosis; Animals; Antibodies, Monoclonal; Biomarkers; CHO Cells; Carbohydrate Sequence; Cell Proliferation; Cricetinae; Cricetulus; Disease Models, Animal; Endothelial Cells; Female; Heparitin Sulfate; Humans; Male; Mice; Neoplasms; Rats; Rats, Wistar; Single-Chain Antibodies; Tumor Necrosis Factor-alpha
Heparan sulfate (HS) is a structurally complex polysaccharide that interacts with a broad spectrum of extracellular effector ligands and thereby is thought to regulate a diverse array of biologic processes. The specificity of HS-ligand interactions is determined by the arrangement of sulfate groups on HS, which creates distinct binding motifs. Biologically important HS motifs are expected to exhibit regulated expression, yet there is a profound lack of tools to identify such motifs; consequently, little is known of their structures and functions. We have identified a novel phage display-derived antibody (NS4F5) that recognizes a highly regulated HS motif (HS(NS4F5)), which we have rigorously identified as (GlcNS6S-IdoA2S)(3). HS(NS4F5) exhibits a restricted expression in healthy adult tissues. Blocking HS(NS4F5) on cells in culture resulted in reduced proliferation and enhanced sensitivity to apoptosis. HS(NS4F5) is up-regulated in tumor endothelial cells, consistent with a role in endothelial cell activation. Indeed, TNF-α stimulated endothelial expression of HS(NS4F5), which contributed to leukocyte adhesion. In a mouse model of severe systemic amyloid protein A amyloidosis, HS(NS4F5) was expressed within amyloid deposits, which were successfully detected by microSPECT imaging using NS4F5 as a molecularly targeted probe. Combined, our results demonstrate that NS4F5 is a powerful tool for elucidating the biological function of HS(NS4F5) and can be exploited as a probe to detect novel polysaccharide biomarkers of disease processes.
Smits, N., Kurup, S., Rops, A., ten Dam, G., Massuger, L., Hafmans, T., Turnbull, J., Spillmann, D., Li, J., Kennel, S., Wall, J., Shworak, N. W., Dekhuijzen, P., van der Vlag, J., & van Kuppevelt, T. (2010). The heparan sulfate motif (GlcNS6S-IdoA2S)3, common in heparin, has a strict topography and is involved in cell behavior and disease.. The Journal of biological chemistry, 285 (52). http://dx.doi.org/10.1074/jbc.M110.153791