Evaluation of fluorescently labeled lectins for noninvasive localization of extracellular polymeric substances in Sphingomonas biofilms
Evaluation of fluorescently labeled lectins for noninvasive localization of extracellular polymeric substances in Sphingomonas biofilms
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DOI：
10.1128/aem.66.8.3487-3491.2000
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发表时间：
2000-08-01
影响因子：
4.4
通讯作者：
Wuertz, S
中科院分区：
文献类型：
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作者：
Johnsen, AR;Hausner, M;Wuertz, S
将3株鞘氨单胞菌培养成生物膜，检测其与5种荧光标记凝集素（Con a型IV-TRITC或-Cy5、Pha-E-TRITC、PNA-TRITC、UEA 1-TRITC和WGA-Texas红）的结合情况。只有ConA和WGA被生物膜显著结合。活凝集素与市售鞘脂单胞外多糖制成的人工生物膜的结合类似于与活生物膜的结合，对活生物膜和人工生物膜的ConA染色可以解释为凝集素与鞘脂分泌的多糖的末端甘露糖基和末端葡萄糖基残基以及鞘脂的末端甘露糖基残基结合。WGA对生物膜的染色只能解释为与鞘脂单胞菌糖脂膜结合、与细胞壁结合或非特异性结合。利用ConA和WGA分别与靶糖葡萄糖和n -乙酰氨基葡萄糖的糖偶联作为凝集素特异性评价的方法；鞘膜单胞菌生物膜和鞘膜单胞菌多糖。我们的研究结果表明，凝集素与生物膜的结合并不一定证明生物膜中细胞外聚合物（EPS）中存在特定的目标糖，凝集素可能与非EPS目标结合或非特异性地粘附在生物膜基质的成分上。
Three strains of Sphingomonas were grown as biofilms and tested for binding of five fluorescently labeled lectins (Con A-type IV-TRITC or -Cy5, Pha-E-TRITC, PNA-TRITC, UEA 1-TRITC, and WGA-Texas red). Only ConA and WGA were significantly bound by the biofilms. Binding of the live lectins to artificial biofilms made of the commercially available Sphinogomonas extracellular polysaccharides was similar to binding to living biofilms, Staining of the living and artificial biofilms by ConA might be explained as binding of the lectin to the terminal mannosyl and terminal glucosyl residues in the polysaccharides secreted by Sphingomonas as well as to the terminal mannosyl residue in glycosphingolipids. Staining of the biofilms by WGA could only be explained as binding to the Sphingomonas glycosphingolipid membrane, binding to the cell wall, or nonspecific binding. Glycoconjugation of ConA and WGA with the target sugars glucose and N-acetylglucosamine, respectively, was used as a method for evaluation of the specificity of the lectin; towards Sphingomonas biofilms and Sphingomonas polysaccharides. Our results show that the binding of lectins to biofilms does not necessarily prove the presence of specific target sugars in the extracellular polymeric substances (EPS) in biofilms, The lectins may bind to non-EPS targets or adhere nonspecifically to components of the biofilm matrix.