Department of Life Sciences, Imperial College London, London SW7 2AZ, United Kingdom.
Department of Pathobiology and Population Sciences, Royal Veterinary College, North Mymms, Hatfield, Hertfordshire AL9 7TA, United Kingdom.
J Biol Chem. 2020 Apr 3;295(14):4541-4555. doi: 10.1074/jbc.RA120.012783. Epub 2020 Feb 24.
Many members of the C-type lectin family of glycan-binding receptors have been ascribed roles in the recognition of microorganisms and serve as key receptors in the innate immune response to pathogens. Other mammalian receptors have become targets through which pathogens enter target cells. These receptor roles have often been documented with binding studies involving individual pairs of receptors and microorganisms. To provide a systematic overview of interactions between microbes and the large complement of C-type lectins, here we developed a lectin array and suitable protocols for labeling of microbes that could be used to probe this array. The array contains C-type lectins from cow, chosen as a model organism of agricultural interest for which the relevant pathogen-receptor interactions have not been previously investigated in detail. Screening with yeast cells and various strains of both Gram-positive and -negative bacteria revealed distinct binding patterns, which in some cases could be explained by binding to lipopolysaccharides or capsular polysaccharides, but in other cases they suggested the presence of novel glycan targets on many of the microorganisms. These results are consistent with interactions previously ascribed to the receptors, but they also highlight binding to additional sugar targets that have not previously been recognized. Our findings indicate that mammalian lectin arrays represent unique discovery tools for identifying both novel ligands and new receptor functions.
许多 C 型凝集素家族的糖结合受体成员被认为在识别微生物方面发挥作用,并作为先天免疫反应中识别病原体的关键受体。其他哺乳动物受体已成为病原体进入靶细胞的靶点。这些受体作用通常通过涉及单个受体对和微生物的结合研究来证明。为了系统地概述微生物与大量 C 型凝集素之间的相互作用,我们在这里开发了一种凝集素阵列和适合标记微生物的协议,可用于探测该阵列。该阵列包含来自奶牛的 C 型凝集素,奶牛被选为农业感兴趣的模式生物,其相关的病原体-受体相互作用以前没有详细研究过。用酵母细胞和革兰氏阳性和阴性细菌的各种菌株进行筛选显示出不同的结合模式,在某些情况下可以通过与脂多糖或荚膜多糖结合来解释,但在其他情况下,这表明许多微生物上存在新的聚糖靶标。这些结果与以前归因于受体的相互作用一致,但它们也突出了与以前未被识别的额外糖靶标的结合。我们的发现表明,哺乳动物凝集素阵列是识别新型配体和新受体功能的独特发现工具。