Microarrays of Synthetic Heparin Oligosaccharides

Abstract

We present the first preparation of microarrays containing synthetic heparin oligosaccharides in order to elucidate the heparin-protein interactions involved in a variety of biological processes. For this purpose, we have developed a novel linker strategy that is compatible with the protecting-group manipulations required for the synthesis of the highly sulfated oligosaccharides and can also be extended to an automated solid phase approach. Strategic placement of the orthogonally protected amine linker was key to the success of the array construction. These heparin chips allow for the high-throughput screening of oligosaccharides by using approximately picomoles of protein. The potential of the new method was demonstrated by probing the carbohydrate affinity of two heparin-binding growth factors, FGF-1 and FGF-2, that are implicated in the development and differentiation of several tumors.

Heparan Sulfate Proteoglycans Containing a Glypican 5 Core and 2-O-Sulfo-iduronic Acid Function as Sonic Hedgehog Co-receptors to Promote Proliferation

“…Microarray Fabrication-Synthetic glycans 1-13 were prepared as described (28,29). Glycan 14 was prepared by functionalization of the deaminated 5-kDa heparin with 1,11-diamino-3,6,9-trioxaundecane by reductive amination.…”

Anticoagulant heparan sulfate: structural specificity and biosynthesis

“…Further, chemical synthesis of non-analog heparin and HS oligosaccharides, larger than hexasaccharides, is extremely difficult, if not impossible, based on currently available methods for carbohydrate synthesis. While a number of groups continue to pursue the synthesis of heparin (48)(49)(50), it is likely that chemical synthesis alone will be incapable of generating most larger oligosaccharide structures. The application of HS biosynthetic enzymes for generating large heparin and HS oligosaccharides with desired biological activities offers a promising alternative approach.…”

Synthetic sugars enhance the functional glycomics toolkit

“…Induced-fit binding energies are often derived from distortion of the ligand and/or receptor, so the solution structures need to be extended to bound conformations with protein partners to be of predictive value. Additionally, further development of glycoarray techniques 2,4,5,8 is clearly necessary. The microarraying and protein-binding interrogation of synthetic CS oligosaccharides reported by Gama et al 2 is similar to that in use for synthetic heparins 5 and illustrates the utility of miniaturized interaction studies.…”

“…Additionally, further development of glycoarray techniques 2,4,5,8 is clearly necessary. The microarraying and protein-binding interrogation of synthetic CS oligosaccharides reported by Gama et al 2 is similar to that in use for synthetic heparins 5 and illustrates the utility of miniaturized interaction studies. However, there remains a need to optimize surface chemistries, coupling efficiencies, and densities, with the goal of producing quantitative data on binding affinities and kinetics.…”

“…Synthesis of iduronate residues is very challenging, yet most known specific GAG-protein interactions involve this residue. Small numbers of synthetic saccharides from the heparan sulfate-heparin family are emerging 5 . There is an evident need for GAG saccharides encompassing a wider range of sulfation and uronate variants, and in longer sequences composed of variant disaccharide building blocks.…”