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Purification and characterization of heparan sulfate from human primary osteoblasts
Author(s) -
Murali Sadasivam,
Manton Kerry J.,
Tjong Vinalia,
Su Xiaodi,
Haupt Larisa M.,
Cool Simon M.,
Nurcombe Victor
Publication year - 2009
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.22340
Subject(s) - heparan sulfate , chemistry , extracellular matrix , biochemistry , cleavage (geology) , glycosaminoglycan , sulfation , affinity chromatography , extracellular , proteoglycan , cell , ligand (biochemistry) , enzyme , biology , receptor , paleontology , fracture (geology)
Heparan sulfate (HS) is a linear, highly variable, highly sulfated glycosaminoglycan sugar whose biological activity largely depends on internal sulfated domains that mediate specific binding to an extensive range of proteins. In this study we employed anion exchange chromatography, molecular sieving and enzymatic cleavage on HS fractions purified from three compartments of cultured osteoblasts—soluble conditioned media, cell surface, and extracellular matrix (ECM). We demonstrate that the composition of HS chains purified from the different compartments is structurally non‐identical by a number of parameters, and that these differences have significant ramifications for their ligand‐binding properties. The HS chains purified of conditioned medium had twice the binding affinity for FGF2 when compared with either cell surface or ECM HS. In contrast, similar binding of BMP2 to the three types of HS was observed. These results suggest that different biological compartments of cultured cells have structurally and functionally distinct HS species that help to modulate the flow of HS‐dependent factors between the ECM and the cell surface. J. Cell. Biochem. 108: 1132–1142, 2009. © 2009 Wiley‐Liss, Inc.