Kuespert Katharina, Hauck Christof R
Department of Biology, University of Konstanz, Konstanz, Germany.
Methods Mol Biol. 2009;470:57-65. doi: 10.1007/978-1-59745-204-5_5.
A critical determinant of host range and specificity relies on the ability of pathogenic bacteria to recognize eukaryotic cell surface molecules via specialized adhesins. The specific adhesin-receptor interaction allows pathogens to tightly bind to their target cells, thereby facilitating the colonization of host tissues. Therefore, the identification and characterization of bacterial adhesins is a major topic in infection biology. This chapter focuses on a rapid and simple method for the analysis of adhesin-receptor interactions that permits the characterization of receptor binding properties at the level of single bacteria. Accordingly, this methodological approach is ideally suited for the analysis of adhesins expressed in a phase-variable manner and for the study of heterogeneous bacterial populations. Besides focusing on the receptor-binding assay, this chapter describes the production of fluorescence-tagged soluble host receptor domains required for conducting this assay.
宿主范围和特异性的一个关键决定因素取决于病原菌通过特殊黏附素识别真核细胞表面分子的能力。特定的黏附素-受体相互作用使病原体能够紧密结合到其靶细胞上，从而促进宿主组织的定殖。因此，细菌黏附素的鉴定和表征是感染生物学中的一个主要课题。本章重点介绍一种快速简便的分析黏附素-受体相互作用的方法，该方法能够在单个细菌水平上表征受体结合特性。因此，这种方法非常适合分析以相变方式表达的黏附素以及研究异质性细菌群体。除了关注受体结合测定外，本章还描述了进行该测定所需的荧光标记可溶性宿主受体结构域的制备。
