Abstract
Glycosylation of bacterial cell surfaces is emerging as a critical factor in symbiosis, pathogenesis, cell-cell interactions and immune evasion1,2,3. The lack of high-throughput analytical tools to examine bacterial glycans has been a major obstacle to the field and has hindered closer examination of the dynamics of carbohydrate variation. We have recently developed a lectin microarray for the analysis of glycoproteins4. Herein we present a rapid analytical system based on this technology for the examination of bacterial glycans. The glycosylation pattern observed distinguishes closely related Escherichia coli strains from one another, providing a facile means of fingerprinting bacteria. In addition, dynamic alterations in the carbohydrate coat of a pathogenic E. coli strain are readily observed. The fast evaluation of real-time alterations in surface-carbohydrate epitopes allows examination of the dynamic role of bacterial sugars in response to external stimuli such as the immune system.
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Acknowledgements
The authors thank D.E. Graham, Department of Chemistry and Biochemistry, University of Texas at Austin, for JM101; S.M. Payne, Department of Molecular Genetics and Microbiology, University of Texas at Austin, for LT2 and HB101; R. Silver and L. Wright, University of Rochester Medical Center, for RS218; I.J. Molineux, Department of Molecular Genetics and Microbiology, University of Texas at Austin; and the Beckman Young Investigator Award for financial support.
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Supplementary information
Supplementary Fig. 1
Growth curve of RS218 in EZ Rich Media. (PDF 24 kb)
Supplementary Fig. 2
Dye uptake as a function of OD600 for cells grown in both EZ Rich Media (Rich Media) and Luria Broth. (PDF 21 kb)
Supplementary Fig. 3
Experimental for 2-color growth-dependent variation of RS218 glycosylation. (PDF 20 kb)
Supplementary Fig. 4
2-color growth dependent variation of RS218 glycosylation (EZ-Rich Defined Media). (PDF 66 kb)
Supplementary Fig. 5
2-color growth dependent variation of RS218 glycosylation (Luria Broth (LB)). (PDF 83 kb)
Supplementary Table 1
Lectin panel. (PDF 23 kb)
Supplementary Table 2
Mean fluorescence and standard deviations. (PDF 10 kb)
Supplementary Table 3
Agglutination data. (PDF 10 kb)
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Hsu, KL., Pilobello, K. & Mahal, L. Analyzing the dynamic bacterial glycome with a lectin microarray approach. Nat Chem Biol 2, 153–157 (2006). https://doi.org/10.1038/nchembio767
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DOI: https://doi.org/10.1038/nchembio767
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