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These small, synthetic, homogeneous molecules bind in the GAG-binding site of proteins resulting in modulation of function.
NSGMs also possesses multiple sulfate groups, in the manner of PI-88, and our earlier work shows that they typically bind in the GAG-binding site on proteins and modulate function.
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This supports our previous findings and indicated that the hMSCs were differentiating towards chondrogenesis in the GAG-binding hydrogels.
In this report, we demonstrate the GAG-binding 'fingerprint' of two important GAG-binding proteins – osteoprotogerin and TIMP-3.
The in silico design process is clearly dependent on the knowledge of the GAG-binding site in the target protein.
(C ) Luciferase reporter assays for Norrin mutations (coloured green) in the GAG binding site.
In combination, these data suggest that the GAG binding site is independent from the Fz4 and putative Lrp5/6 binding sites.
Engineering only the hydrophobic pocket in the proximal loop of the chemokine did not increase the GAG binding.
For the sake of higher biological relevance, we decided to carry out the GAG binding measurements with hexameric GAGs.
In our original manuscript we reported that mutations (R107E/R109E/R115L) in the Norrin GAG binding site do not affect the ability of Norrin to bind Fz4CRD but do impair the activation of signalling.
In order to explore the effect of GAG elongation on binding to IL-8, we used 8-, 10- and 12-mers of HA463′ bound in the common binding pose predicted for tetrameric and hexameric GAGs to have high affinity.
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