Department of Infectious Diseases and Immunology, Utrecht University, Utrecht, the Netherlands.
Cell Microbiol. 2009 Dec;11(12):1768-81. doi: 10.1111/j.1462-5822.2009.01370.x. Epub 2009 Aug 13.
An increasing number of bacterial pathogens produce an array of glycoproteins of unknown function. Here we report that Campylobacter jejuni proteins that are modified by the N-linked glycosylation machinery encoded by the pgl locus bind the human Macrophage Galactose-type lectin (MGL). MGL receptor binding was abrogated by EDTA and N-acetylgalactosamine (GalNAc) and was successfully transferred to Escherichia coli by introducing the C. jejuni pgl locus together with a glycan acceptor protein. In addition to glycoproteins, C. jejuni lipooligosaccharide with a terminal GalNAc residue was recognized by MGL. Recombinant E. coli expressing the C. jejuni pgl locus in the absence of a suitable glycan acceptor protein produced altered lipopolysaccharide glycoforms that gained MGL reactivity. Infection assays demonstrated high levels of GalNAc-dependent interaction of the recombinant E. coli with MGL-transfected mammalian cells. In addition, interleukin-6 production by human dendritic cells was enhanced by C. jejuni lacking N-linked glycans compared with wild-type bacteria. Collectively, our results provide evidence that both N-linked glycoproteins and distinct lipooligosaccharide glycoforms of C. jejuni are ligands for the human C-type lectin MGL and that the C. jejuni N-glycosylation machinery can be exploited to target recombinant bacteria to MGL-expressing eukaryotic cells.
越来越多的细菌病原体产生了一系列具有未知功能的糖蛋白。在这里,我们报告称,由 pgl 基因座编码的 N-连接糖基化机制修饰的空肠弯曲菌蛋白与人类巨噬细胞半乳糖型凝集素(MGL)结合。MGL 受体结合被 EDTA 和 N-乙酰半乳糖胺(GalNAc)阻断,并通过引入空肠弯曲菌 pgl 基因座和糖受体蛋白成功转移到大肠杆菌中。除糖蛋白外,带有末端 GalNAc 残基的空肠弯曲菌脂寡糖也被 MGL 识别。在没有合适糖受体蛋白的情况下表达空肠弯曲菌 pgl 基因座的重组大肠杆菌产生了改变的脂多糖糖型,获得了 MGL 反应性。感染实验表明,重组大肠杆菌与转染了 MGL 的哺乳动物细胞之间存在高水平的依赖于 GalNAc 的相互作用。此外,与野生型细菌相比,缺乏 N-连接聚糖的空肠弯曲菌可增强人树突状细胞产生的白细胞介素 6。总的来说,我们的结果提供了证据,证明空肠弯曲菌的 N-连接糖蛋白和不同的脂寡糖糖型都是人类 C 型凝集素 MGL 的配体,并且空肠弯曲菌的 N-糖基化机制可被利用来将重组细菌靶向表达 MGL 的真核细胞。