Zhu X-Y, Holtz Bryan, Wang Yini, Wang Lai-Xi, Orndorff Paul E, Guo Athena
Department of Chemistry, University of Minnesota, Minneapolis, Minnesota 55455, USA.
J Am Chem Soc. 2009 Sep 30;131(38):13646-50. doi: 10.1021/ja902783n.
A hallmark of cell-surface processes involving glycans is their multivalent interaction with glycan binding proteins (GBPs). Such a multivalent interaction depends critically on the mobility and density of signaling molecules on the membrane surface. While glycan microarrays have been used in exploring multivalent interactions, the lack of mobility and the difficulty in controlling surface density both limit their quantitative applications. Here we apply a fluidic glycan microarray, with glycan density varying for orders of magnitude, to profile cell surface interaction using a model system, the adhesion of Escherichia coli to mannose. We show the quantitative determination of monovalent and multivalent adhesion channels; the latter can be inhibited by nanopartices presenting a high density of mannosyl groups. These results reveal a new E. coli adhesion mechanism: the switching in the FimH adhesion protein avidity from monovalent to multivalent as the density of mobile mannosyl groups increases; such avidity switching enhances binding affinity and triggers multiple fimbriae anchoring. Affinity enhancement toward FimH has only been observed before for oligo-mannose due to the turn on of secondary interactions outside the mannose binding pocket. We suggest that the new mechanism revealed by the fluidic microarray is of general significance to cell surface interactions: the dynamic clustering of simple sugar groups (homogeneous or heterogeneous) on the fluidic membrane surface may simulate the functions of complex glycan molecules.
涉及聚糖的细胞表面过程的一个标志是它们与聚糖结合蛋白(GBP)的多价相互作用。这种多价相互作用严重依赖于膜表面信号分子的流动性和密度。虽然聚糖微阵列已被用于探索多价相互作用,但缺乏流动性以及控制表面密度的困难都限制了它们的定量应用。在这里,我们应用一种流体聚糖微阵列,其聚糖密度在几个数量级上变化,使用一个模型系统——大肠杆菌与甘露糖的黏附——来描绘细胞表面相互作用。我们展示了单价和多价黏附通道的定量测定;后者可被呈现高密度甘露糖基的纳米颗粒抑制。这些结果揭示了一种新的大肠杆菌黏附机制:随着可移动甘露糖基密度的增加,FimH黏附蛋白的亲和力从单价转变为多价;这种亲和力转换增强了结合亲和力并触发多个菌毛锚定。以前仅在寡聚甘露糖中观察到对FimH亲和力的增强,这是由于甘露糖结合口袋外二级相互作用的开启。我们认为,流体微阵列揭示的新机制对细胞表面相互作用具有普遍意义:流体膜表面上简单糖基(同质或异质)的动态聚集可能模拟复杂聚糖分子的功能。