Purification and characterization of heparan sulfate from human primary osteoblasts
Murali, Sadasivam , Manton, Kerry J., Tjong, Vinalia , Su, Xiaodi , Haupt, Larisa M., Cool, Simon M. , & Nurcombe, Victor (2009) Purification and characterization of heparan sulfate from human primary osteoblasts. Journal of Cellular Biochemistry, 108(5), pp. 1132-1142.
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.
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|Item Type:||Journal Article|
|Keywords:||glycosaminoglycans, extracellular matrix, growth factors, bone, differentiation|
|Subjects:||Australian and New Zealand Standard Research Classification > BIOLOGICAL SCIENCES (060000) > BIOCHEMISTRY AND CELL BIOLOGY (060100)|
Australian and New Zealand Standard Research Classification > BIOLOGICAL SCIENCES (060000) > BIOCHEMISTRY AND CELL BIOLOGY (060100) > Cell Development Proliferation and Death (060103)
|Divisions:||Current > Schools > School of Biomedical Sciences|
Current > QUT Faculties and Divisions > Faculty of Health
Current > Institutes > Institute of Health and Biomedical Innovation
|Deposited On:||25 Aug 2011 08:17|
|Last Modified:||07 Apr 2014 09:00|
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