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A lectin microarray approach for the rapid analysis of bacterial glycans

Nature Protocols volume 1, pages 543–549 (2006)Cite this article

Abstract

Rapid evaluation of microbial cell-surface carbohydrates is essential to understanding the mechanisms by which bacteria use glycans to establish pathogenic or symbiotic relationships. Microbial glycan analysis is complicated both by the vast diversity of possible carbohydrate structures and by their dynamic nature. Bacteria can rapidly alter their glycan coats by switching the genes that are involved on and off in a phase-variable manner. Currently, there is a lack of appropriate tools for studying dynamic carbohydrate alterations. Here, we present a lectin microarray protocol for the high-throughput evaluation of cell-surface microbial sugars. The binding patterns of fluorescent bacteria to these arrays provide a simple means to fingerprint bacteria based on their surface carbohydrates. In addition, this method provides a rapid, parallel evaluation of glycans from multiple bacterial samples, allowing dynamic changes in carbohydrate structures to be studied. The entire procedure takes ∼12 h but the printing of the microarray can be performed in advance.

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Figure 1: A flow diagram of the lectin microarray protocol.
Figure 2: Titration curve for uptake of SYTO 85 dye by Escherichia coli strains JM101 and HB101.
Figure 3: Lectin-binding patterns of Escherichia coli HB101, representative of a minimum of three replicates.

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Acknowledgements

The authors thank D.E. Graham, Department of Chemistry and Biochemistry, University of Texas at Austin for JM101 and S.M. Payne, Department of Molecular Genetics and Microbiology, University of Texas at Austin for HB101 and the Beckman Young Investigator Award for Financial Support.

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Authors and Affiliations

  1. Department of Chemistry and Biochemistry, University of Texas at Austin, 1 University Station, A5300, Austin, 78712-0265, Texas, USA

    Ku-Lung Hsu

  2. Center for Systems and Synthetic Biology and Institute for Cellular and Molecular Biology, University of Texas at Austin, 1 University Station, A5300, Austin, 78712-0265, Texas, USA

    Lara K Mahal

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  1. Ku-Lung Hsu
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Correspondence to Lara K Mahal.

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Hsu, KL., Mahal, L. A lectin microarray approach for the rapid analysis of bacterial glycans. Nat Protoc 1, 543–549 (2006). https://doi.org/10.1038/nprot.2006.76

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