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Analyzing the dynamic bacterial glycome with a lectin microarray approach


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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Figure 1: Lectin array.
Figure 2: Glycopatterns of pathogenic and nonpathogenic bacterial strains.
Figure 3: Carbohydrate inhibition.
Figure 4: Growth-dependent variation of neonatal meningitis E. coli strain RS218 glycosylation.

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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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Correspondence to Lara K Mahal.

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The authors declare no competing financial interests.

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).

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