An engineered mannoside presenting platform:: Escherichia coli adhesion under static and dynamic conditions
An engineered mannoside presenting platform:: Escherichia coli adhesion under static and dynamic conditions
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DOI：
10.1002/adfm.200701246
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发表时间：
2008-05-09
影响因子：
19
通讯作者：
Textor, Marcus
中科院分区：
文献类型：
--
作者：
Barth, Katrin A.;Coullerez, Geraldine;Textor, Marcus
病原体对宿主组织的粘附机制的研究已经获得了越来越多的兴趣，因为细菌粘附参与了表面定殖和感染的早期阶段。在这里，我们描述了一个平台，研究细菌大肠杆菌（E。大肠杆菌）K-12菌株与模拟细胞界面的分子定义明确的表面。该方法使用聚（乙二醇）刷界面，其显示高甘露糖N-连接聚糖的合成决定簇，密度范围为（3.8 x 10（4）-1.6 x 10（5）甘露糖苷μ m（-2）），用于研究与细菌的多价相互作用。细菌附着是由位于细菌I型皮利顶端的粘附蛋白FimH和甘露糖基化表面之间的特异性相互作用介导的。利用合成工程甘露糖，发现强粘附细菌的数量由许多结构物理参数共同调节。除了对碳水化合物密度的依赖外，E.与单甘露糖相比，大肠杆菌中的寡甘露糖与支链三甘露糖Man（α 1 - 3）（Man（α 1 - 6））Man结合，而在其非还原端暴露Man（α 1 - 2）Man的较大寡甘露糖显示低结合能力。甘露糖部分和PEG之间使用的接头也影响E.杆菌与（亲水性）三（EG）相比，（疏水性）丙基接头导致更高的细菌数量，这可能是通过疏水相互作用额外稳定结合复合物的结果。此外，在停滞和流动条件之间的细菌附着中观察到差异，这取决于甘露糖配体的类型。最后，抗微生物剥离与糖基共聚物的位点选择性组装相结合，用于产生微图案，其中细菌菌落被限制在限定的区域和受控的菌落数。
The study of the adhesion mechanisms of pathogens to host tissues has gained increased interest as bacterial adhesion is involved in the early stages of surface colonization and infection. Here we describe a platform to study the specific binding of the bacterium Escherichia coli (E. coli) K-12 strain to molecularly well-defined surfaces mimicking cellular interfaces. This approach uses a poly(ethylene glycol) brush interface, which displays synthetic determinants of the high mannose N-linked glycans in a range of densities (3.8 x 10(4)-1.6 x 10(5) mannosides mu m(-2)) for the investigation of multivalent interactions with bacteria. The bacterial attachment is mediated by specific interactions between the adhesive protein FimH located on the tip of the bacterial type I pili and the mannosylated surfaces. With synthetically engineered mannoses, it is found that the number of strongly adhering bacteria is co-regulated by many structural physical parameters. Beyond the dependency on carbohydrate density, higher numbers of E. coli attach to the branched trimannose Man(alpha 1-3)(Man(alpha 1-6))Man compared to the monomannose, while larger oligomannoses exposing Man(alpha 1-2) Man at their non reducing end show low binding capacity. The linker used between the mannose moiety and PEG is also affecting the binding efficacy of E. coli. The (hydrophobic) propyl linker results in higher bacteria numbers in comparison to the (hydrophilic) tri(EG), likely a consequence of additional stabilization of the binding complex by hydrophobic interactions. Furthermore, differences are observed in bacteria attachment between stagnant and flow conditions that depend on the type of mannose ligand. Finally, a photolithographic resist lift-off combined with site-selective assembly of the glycopolymers is used to produce micropatterns with bacteria colonies confined to defined areas and at controlled colony numbers.