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Oxidative release of natural glycans for functional glycomics

Nature Methods volume 13, pages 528–534 (2016)Cite this article

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Abstract

Glycans have essential roles in biology and the etiology of many diseases. A major hurdle in studying glycans through functional glycomics is the lack of methods to release glycans from diverse types of biological samples. Here we describe an oxidative strategy using household bleach to release all types of free reducing N-glycans and O-glycan-acids from glycoproteins, and glycan nitriles from glycosphingolipids. Released glycans are directly useful in glycomic analyses and can be derivatized fluorescently for functional glycomics. This chemical method overcomes the limitations in glycan generation and promotes archiving and characterization of human and animal glycomes and their functions.

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Figure 1: Comparison of ORNG by NaClO with traditional glycomics methods.
Figure 2: NaClO treatment of glycoproteins to release N-glycans.
Figure 3: Lectin binding on the microarray of N-glycan–AEAB conjugates.
Figure 4: NaClO treatment of glycoproteins to release O-glycans.
Figure 5: Release and tagging of glycans from GSLs by NaClO.

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Acknowledgements

We thank J. Heimburg-Molinaro (Beth Israel Deaconess Medical Center) for help in scientific discussion and preparing the manuscript. We thank H. Sun (Wuhan University) for providing rAAL2 lectin. This work was supported by the US National Institutes of Health (EUREKA grant GM085448 to D.F.S., BTRC grant P41GM10369 to R.D.C. and Common Fund Glycoscience grant U01GM116254 to X.S.), the US Department of Health and Human Services contract HHSN272201400004C (NIAID Centers of Excellence for Influenza Research and Surveillance) and the Defense Advanced Research Projects Agency (Grant HR0011-10-00 to R.D.C.).

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Authors and Affiliations

  1. Department of Biochemistry, Emory University School of Medicine, Atlanta, Georgia, USA

    Xuezheng Song, Hong Ju, Yi Lasanajak, Matthew R Kudelka & David F Smith

  2. Department of Surgery, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts, USA

    Richard D Cummings

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  1. Xuezheng Song
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Contributions

X.S., D.F.S. and R.D.C. conceived the method; X.S., H.J., M.R.K. and Y.L. performed experiments; X.S., D.F.S. and R.D.C. analysed the data and wrote the paper.

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Correspondence to Xuezheng Song or Richard D Cummings.

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Song, X., Ju, H., Lasanajak, Y. et al. Oxidative release of natural glycans for functional glycomics. Nat Methods 13, 528–534 (2016). https://doi.org/10.1038/nmeth.3861

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