Glycan conjugates are an essential part of cell membranes. Each organism possesses a unique glycan conjugate signature that is thus an ideal target for the development of organism specific detection tools.
A number of Clickable Azide-functionalized saccharides such as Kdo-Azide, 6-Azido-Trehalose pLeg-Azide are now available for non-radioactive metabolic labeling of bacterial cell membrane (Fig. 1, Tab. 1). They are cell-permeable and biosynthetically processed instead of their natural counterparts.
The resulting Azide-functionalized bacterial cell membrane can subsequently be visualized via Cu(I)-catalyzed Alkyne-Azide cycloaddition (CuAAC) or Cu(I)-free strain-promoted Alkyne-Azide cycloaddition (SPAAC) that allows to introduce
- a (Desthio)Biotin group for subsequent purification (via Alkynes of (Desthio)Biotin or DBCO-containing (Desthio)Biotin) or
- a fluorescent group for subsequent microscopic imaging (via fluorescent Alkynes or DBCO-containing fluorescent dyes)
Figure 1: CLICK-labeling of bacterial cell membrane is performed by a two-step procedure. 1) Azide-functionalization of bacterial cell membrane by metabolic incorporation of correspondingly-functionalized saccharides and 2) CLICK-labeling via Alkyne- or DBCO-functionalized detection molecules by CuAAC or SPAAC, respectively.
Table 1: Available Azide-functionalized saccharides for bacterial cell membrane labeling.
|Tested bacterial strains
Escherichia coli O86
Escherichia coli K12
||Mycobacterium smegmatis mc2155
Mycobacterium tuberculosis H37Rv
Mycobacterium bovis BCG
||Legionella pneumophila SG1 (Paris), SG1 (Lens), SG1 (Philadelphia), SG3, SG4, SG5, SG6
Also check out our viral and microbial native antigens and recombinant proteins!
 Singer et al. (1972) The fluid mosaic model of the structure of cell membranes. Science 175:720.
 Dumont et al. (2012) Click-Mediated Labeling of Bacterial Membranes through Metabolic Modification of the Lipopolysaccharide Inner Core. Angew. Chem. Int. Ed. 51:3143.
 Swarts et al. (2012) Probing the Myobacterial Trehalome with Bioorthogonal Chemistry. J. Am. Chem. Soc. 134 (39):16123.
 Mas Pons et al. (2014) Identification of Living Legionella pneumophila using species-specific metabolic lipopolysaccharide Labeling. Angew. Chem. Int. Ed. 53:1275.