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Interactions between Sclerostin and Glycosaminoglycans

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Abstract

Sclerostin (SOST) is a glycoprotein having many important functions in the regulation of bone formation as a key negative regulator of Wnt signaling in bone. Surface plasmon resonance (SPR), which allows for a direct quantitative analysis of the label-free molecular interactions in real-time, has been widely used for the biophysical characterization of glycosaminoglycan (GAG)-protein interactions. In the present study, we report kinetics, structural analysis and the effects of physiological conditions (e.g., salt concentrations, Ca2+ and Zn2+concentrations) on the interactions between GAGs and recombinant human (rh) and recombinant mouse (rm) SOST using SPR. SPR results revealed that both SOSTs bind heparin with high affinity (rhSOST-heparin, KD~36 nM and rmSOST-heparin, KD~77 nM) and the shortest oligosaccharide of heparin that effectively competes with full size heparin for SOST binding is octadecasaccharide (18mer). This heparin binding protein also interacts with other highly sulfated GAGs including, disulfated-dermatan sulfate and chondroitin sulfate E. In addition, liquid chromatography-mass spectrometry was used to characterize the structure of sulfated GAGs that bound to SOST.

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Abbreviations

SOST:

Sclerostin

GAG:

Glycosaminoglycan

rhSOST:

Recombinant human sclerostin

rmSOST:

Recombinant mouse sclerostin

SPR:

Surface plasmon resonance

HS:

Heparan sulfate

CSA:

Chondroitin sulfate A

CSB:

Chondroitin sulfate B

DS:

Dermatan sulfate

CSC:

Chondroitin sulfate C

CSD:

Chondroitin sulfate D

CSE:

Chondroitin sulfate E

LMWH:

Low molecular weight heparin

SA:

Streptavidin

dp:

Degree of polymerization

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Acknowledgements

This work was supported by National Institutes of Health Grants: DK111958, CA231074 and AG062344 to R.J.L.

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Authors and Affiliations

  1. Department of Chemical and Biological Engineering, Rensselaer Polytechnic Institute, Troy, NY, 12180, USA

    Fuming Zhang & Robert J. Linhardt

  2. Department of Chemistry and Chemical Biology, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY, 12180, USA

    Jing Zhao, Xinyue Liu & Robert J. Linhardt

  3. Departments of Biological Sciences and Biomedical Engineering, Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY, 12180, USA

    Robert J. Linhardt

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  1. Fuming Zhang
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Correspondence to Fuming Zhang or Robert J. Linhardt.

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Zhang, F., Zhao, J., Liu, X. et al. Interactions between Sclerostin and Glycosaminoglycans. Glycoconj J 37, 119–128 (2020). https://doi.org/10.1007/s10719-019-09900-3

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