Functional Interaction Analysis of GM1-Related Carbohydrates and Vibrio cholerae Toxins Using Carbohydrate Microarray
- Functional Interaction Analysis of GM1-Related Carbohydrates and Vibrio cholerae Toxins Using Carbohydrate Microarray
- 서정현; 김창섭; 차형준[차형준]
- SURFACE-PLASMON RESONANCE; ESCHERICHIA-COLI; LIGAND-BINDING; FLOW-CYTOMETRY; ENTERO-TOXIN; PROTEIN; GM1; OLIGOSACCHARIDE; SPECIFICITY; RECEPTOR
- Issue Date
- AMER CHEMICAL SOC
- ANALYTICAL CHEMISTRY, v.84, no.15, pp.6884 - 6890
- The development of analytical tools is important for understanding the infection mechanisms of pathogenic bacteria or viruses. In the present work, a functional carbohydrate microarray combined with a fluorescence immunoassay was developed to analyze the interactions of Vibrio cholerae toxin (ctx) proteins and GM1-related carbohydrates. Ctx proteins were loaded onto the surface-immobilized GM1 pentasaccharide and six related carbohydrates, and their binding affinities were detected immunologically. The analysis of the ctx-carbohydrate interactions revealed that the intrinsic selectivity of do( was GM1 pentasaccharide >> GM2 tetrasaccharide > asialo >= GM1 tetrasaccharide >= GM3trisaccharide, indicating that a two-finger grip formation and the terminal monosaccharides play important roles in the ctx-GM1 interaction. In addition, whole cholera toxin (ctxAB(5)) had a stricter substrate specificity and a stronger binding affinity than only the cholera toxin B subunit (ctxB). On the basis of the quantitative analysis, the carbohydrate microarray showed the sensitivity of detection of the ctxAB(5)-GM1 interaction with a limit-of-detection (LOD) of 2 ng mL(-1) (23 pM), which is comparable to other reported high sensitivity assay tools. In addition, the carbohydrate microarray successfully detected the actual toxin directly secreted from V. cholerae, without showing cross-reactivity to other bacteria. Collectively, these results demonstrate that the functional carbohydrate microarray is suitable for analyzing toxin protein-carbohydrate interactions and can be applied as a biosensor for toxin detection.
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