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Metabolic installation of thiols into sialic acid modulates adhesion and stem cell biology

Nature Chemical Biology volume 2, pages 149–152 (2006)Cite this article

Abstract

Metabolic 'oligosaccharide engineering' methods based on N-acetyl-D-mannosamine (ManNAc) analogs allow the glycocalyx of living cells to be remodeled1,2,3. Herein we report the analog Ac5ManNTGc (1) that enables thiols to be expressed in surface sialic acids. By locating this versatile functional group on the outer periphery of normally nonadhesive human Jurkat cells, we obtained spontaneous cell-cell clustering and attachment to complementary maleimide-derivatized substrates. When analyzed in human embryoid body–derived (hEBD) stem cells, Ac5ManNTGc induced β-catenin expression and altered cell morphology, consistent with neuronal differentiation. Notably, these effects were modulated by the growth substrate of the cells, with a stronger response observed on a gold surface than on glass. Together, these results establish sugar analogs as small-molecule tools for tissue engineering by providing a method for attaching cells to scaffolds via their surface carbohydrates as well as offering a means to influence stem cell fates.

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Figure 1: Surface display of thiol-bearing sialic acids.
Figure 2: Analog-mediated cell clustering and attachment to complementary surfaces.
Figure 3: Metabolic incorporation of Ac5ManNTGc (1) in hEBD-LVEC cells.
Figure 4: Analogs alter the morphology of hEBD-LVEC cells.
Figure 5: Upregulation of β-catenin in hEBD-LVEC cells induced by 1.

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Acknowledgements

We are grateful to K. Konstantopoulos for flow cytometer access, P. Pawar for technical assistance in FACS experiments, J.D. Gearhart and M.J. Shamblott for the kind gift of EBD LVEC cells, T.H. Wang for the kind gift of QD655-streptavidin conjugate, and J.M. McCaffery and E. Perkins of the Integrated Imaging Facility (Johns Hopkins University Department of Biology) for help with confocal microscopy. This work was supported by funding from the Arnold and Mabel Beckman Foundation, the US National Institutes of Health (1R01CA112314-01A1) and the National Science Foundation (QSB-0425668).

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  1. Department of Biomedical Engineering, The Johns Hopkins University, 3400 N. Charles Street, Clark Hall 106A, Baltimore, 21218, Maryland, USA

    Srinivasa-Gopalan Sampathkumar, Adrienne V Li, Mark B Jones, Zhonghui Sun & Kevin J Yarema

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  1. Srinivasa-Gopalan Sampathkumar
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Correspondence to Kevin J Yarema.

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Supplementary information

Supplementary Fig. 1

Concentration and time course of CST expression. (PDF 72 kb)

Supplementary Fig. 2

Dose dependency of CST expression on 1. (PDF 230 kb)

Supplementary Scheme 1

Synthesis of 1. (PDF 52 kb)

Supplementary Scheme 2

Synthesis of 2. (PDF 50 kb)

Supplementary Scheme 3

Labeling of cell-surface thiols. (PDF 51 kb)

Supplementary Scheme 4

Effect of TCEP treatment. (PDF 43 kb)

Supplementary Methods (PDF 488 kb)

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Sampathkumar, SG., Li, A., Jones, M. et al. Metabolic installation of thiols into sialic acid modulates adhesion and stem cell biology. Nat Chem Biol 2, 149–152 (2006). https://doi.org/10.1038/nchembio770

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