title: The heparan sulfate motif (GlcNS6S-IdoA2S)3, common in heparin, has a strict topography and is involved in cell behavior and disease.
abstract: 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.
authors.affiliation: Department of Biochemistry, Radboud University Nijmegen Medical Centre, Nijmegen Centre for Molecular Life Sciences, 6500 HB Nijmegen, The Netherlands.
authors.forename: Nicole C
authors.initials: NC
authors.lastname: Smits
authors.forename: Sindhulakshmi
authors.initials: S
authors.lastname: Kurup
authors.forename: Angelique L
authors.initials: AL
authors.lastname: Rops
authors.forename: Gerdy B
authors.initials: GB
authors.lastname: ten Dam
authors.forename: Leon F
authors.initials: LF
authors.lastname: Massuger
authors.forename: Theo
authors.initials: T
authors.lastname: Hafmans
authors.forename: Jeremy E
authors.initials: JE
authors.lastname: Turnbull
authors.forename: Dorothe
authors.initials: D
authors.lastname: Spillmann
authors.forename: Jin-ping
authors.initials: JP
authors.lastname: Li
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Kennel
authors.forename: Jonathan S
authors.initials: JS
authors.lastname: Wall
authors.forename: Nicholas W
authors.initials: NW
authors.lastname: Shworak
authors.forename: P N Richard
authors.initials: PN
authors.lastname: Dekhuijzen
authors.forename: Johan
authors.initials: J
authors.lastname: van der Vlag
authors.forename: Toin H
authors.initials: TH
authors.lastname: van Kuppevelt
mesh_terms: D058227:Amyloidogenic Proteins / Q000276:immunology / Q000378:metabolism; D000686:Amyloidosis / Q000276:immunology / Q000378:metabolism*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000494:pharmacology*; D015415:Biomarkers / Q000378:metabolism; D016466:CHO Cells; D002240:Carbohydrate Sequence; D049109:Cell Proliferation / Q000187:drug effects; D006224:Cricetinae; D003412:Cricetulus; D004195:Disease Models, Animal; D042783:Endothelial Cells / Q000276:immunology / Q000378:metabolism*; D005260:Female; D006497:Heparan Sulfate / Q000037:antagonists & inhibitors / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D008297:Male; D051379:Mice; D009369:Neoplasms / Q000276:immunology / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D057127:Single-Chain Antibodies / Q000276:immunology / Q000494:pharmacology*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
chemical_list: D058227:Amyloidogenic Proteins; D000911:Antibodies, Monoclonal; D015415:Biomarkers; D006497:Heparan Sulfate; D057127:Single-Chain Antibodies; D014409:Tumor Necrosis Factor-alpha