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Sulfation patterns of glycosaminoglycans encode molecular recognition and activity

Nature Chemical Biology volume 2, pages 467–473 (2006)Cite this article

Abstract

Although glycosaminoglycans contribute to diverse physiological processes1,2,3,4, an understanding of their molecular mechanisms has been hampered by the inability to access homogeneous glycosaminoglycan structures. Here, we assembled well-defined chondroitin sulfate oligosaccharides using a convergent, synthetic approach that permits installation of sulfate groups at precise positions along the carbohydrate backbone. Using these defined structures, we demonstrate that specific sulfation motifs function as molecular recognition elements for growth factors and modulate neuronal growth. These results provide both fundamental insights into the role of sulfation and direct evidence for a 'sulfation code' whereby glycosaminoglycans encode functional information in a sequence-specific manner analogous to that of DNA, RNA and proteins.

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Figure 1: A specific sulfation pattern promotes the interaction of CS with neuronal growth factors.
Figure 2: The sulfation pattern directs the neuritogenic activity of CS.
Figure 3: The CS-E sulfation motif stimulates neuronal growth through activation of the midkine-PTPζ and BDNF-TrkB signaling pathways.

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Acknowledgements

We thank V.W.T. Kam for assistance with modifying the Dreiding force field; R.H. Grubbs for the catalyst; C.J. Rogers for assistance with data analysis; J.L. Riechmann, director of the Millard and Muriel Jacobs Genetic and Genomics Laboratory at Caltech, for assistance with printing the microarrays; S. Ou in the Caltech Monoclonal Antibody Facility; and the Biological Imaging Center and Environmental Analysis Center at Caltech for instrumentation. This work was supported by the American Cancer Society (RSG-05-106-01-CDD), Human Frontiers Science Program, Tobacco-Related Disease Research Program (14RT-0034), National Institutes of Health (RO1 NS045061, C.I.G. and L.C.H.-W.), National Science Foundation (S.E.T.) and Howard Hughes Medical Institute.

Author information

Author notes

  1. Nagarajan Vaidehi

    Present address: Division of Immunology, City of Hope Cancer Research Center, Beckman Research Institute of City of Hope, 1500 East Duarte Road, Duarte, California, 91010, USA

  2. Cristal I Gama and Sarah E Tully: These authors contributed equally to this work.

Authors and Affiliations

  1. Division of Chemistry and Chemical Engineering and Howard Hughes Medical Institute, California Institute of Technology, Pasadena, 91125, California, USA

    Cristal I Gama, Sarah E Tully, Peter M Clark, Manish Rawat & Linda C Hsieh-Wilson

  2. Department of Pharmacology, Kurume University School of Medicine, 67 Asahi-machi, Kurume, 830-0011, Fukuoka, Japan

    Naoki Sotogaku & Akinori Nishi

  3. Materials and Process Simulation Center (MSC), California Institute of Technology, Pasadena, 91125, California, USA

    Nagarajan Vaidehi & William A Goddard III

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C.I.G. and S.E.T. contributed equally to this work.

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Correspondence to Linda C Hsieh-Wilson.

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Supplementary information

Supplementary Fig. 1

Each CS tetrasaccharide favors a distinct set of torsion angles: torsion plots for ψ1 and φ1. (PDF 656 kb)

Supplementary Fig. 2

Each CS tetrasaccharide favors a distinct set of torsion angles: torsion plots for ψ2 and φ2. (PDF 586 kb)

Supplementary Fig. 3

Each CS tetrasaccharide favors a distinct set of torsion angles: torsion plots for ψ3 and φ3. (PDF 587 kb)

Supplementary Fig. 4

Each CS tetrasaccharide presents a unique van der Waals surface. (PDF 35 kb)

Supplementary Fig. 5

Monoclonal antibody 5D2-2C2 (raised against CS-C) selectively recognizes the CS-C tetrasaccharide. (PDF 44 kb)

Supplementary Fig. 6

Antibodies against BDNF or midkine, but not control IgG antibodies, block the binding of BDNF or midkine to CS-E on the microarray. (PDF 49 kb)

Supplementary Fig. 7

Function-blocking antibodies against BDNF and TrkB, but not class-matched control antibodies, disrupt the neuritogenic activity of CS-E. (PDF 47 kb)

Supplementary Methods

Experimental procedures for the synthesis of CS tetrasaccharides 1, 2, 3 and 4, conjugation methods for preparing carbohydrate microarrays, and dot blot procedures. (PDF 380 kb)

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Gama, C., Tully, S., Sotogaku, N. et al. Sulfation patterns of glycosaminoglycans encode molecular recognition and activity. Nat Chem Biol 2, 467–473 (2006). https://doi.org/10.1038/nchembio810

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