Department of Microbiology and Molecular Medicine, Faculty of Medicine, University of Geneva Centre Medical Universitaire, 1 Rue Michel-Servet, 1211 Geneva 4, Switzerland.
J Biol Chem. 2010 Jan 15;285(3):2064-76. doi: 10.1074/jbc.M109.060988. Epub 2009 Nov 9.
Numerous intracellular pathogens exploit cell surface glycoconjugates for host cell recognition and entry. Unlike bacteria and viruses, Toxoplasma gondii and other parasites of the phylum Apicomplexa actively invade host cells, and this process critically depends on adhesins (microneme proteins) released onto the parasite surface from intracellular organelles called micronemes (MIC). The microneme adhesive repeat (MAR) domain of T. gondii MIC1 (TgMIC1) recognizes sialic acid (Sia), a key determinant on the host cell surface for invasion by this pathogen. By complementation and invasion assays, we demonstrate that TgMIC1 is one important player in Sia-dependent invasion and that another novel Sia-binding lectin, designated TgMIC13, is also involved. Using BLAST searches, we identify a family of MAR-containing proteins in enteroparasitic coccidians, a subclass of apicomplexans, including T. gondii, suggesting that all these parasites exploit sialylated glycoconjugates on host cells as determinants for enteric invasion. Furthermore, this protein family might provide a basis for the broad host cell range observed for coccidians that form tissue cysts during chronic infection. Carbohydrate microarray analyses, corroborated by structural considerations, show that TgMIC13, TgMIC1, and its homologue Neospora caninum MIC1 (NcMIC1) share a preference for alpha2-3- over alpha2-6-linked sialyl-N-acetyllactosamine sequences. However, the three lectins also display differences in binding preferences. Intense binding of TgMIC13 to alpha2-9-linked disialyl sequence reported on embryonal cells and relatively strong binding to 4-O-acetylated-Sia found on gut epithelium and binding of NcMIC1 to 6'sulfo-sialyl Lewis(x) might have implications for tissue tropism.
许多细胞内病原体利用细胞表面糖缀合物来识别和进入宿主细胞。与细菌和病毒不同,刚地弓形虫和其他顶复门寄生虫主动入侵宿主细胞,这一过程严重依赖于从称为微线体(MIC)的细胞内细胞器释放到寄生虫表面的黏附素(微线体蛋白)。刚地弓形虫 MIC1(TgMIC1)的微线体黏附重复(MAR)结构域识别唾液酸(Sia),这是该病原体入侵宿主细胞表面的关键决定因素。通过互补和入侵实验,我们证明 TgMIC1 是依赖 Sia 的入侵的重要参与者,另一种新型 Sia 结合凝集素,称为 TgMIC13,也参与其中。使用 BLAST 搜索,我们在肠内寄生的球虫类(顶复门的一个亚类)中发现了一个含有 MAR 的蛋白质家族,包括刚地弓形虫,这表明所有这些寄生虫都将宿主细胞上的唾液酸化糖缀合物用作肠道入侵的决定因素。此外,该蛋白质家族可能为观察到的广泛宿主细胞范围提供了基础,这些寄生虫在慢性感染期间形成组织囊肿。碳水化合物微阵列分析,结合结构考虑,表明 TgMIC13、TgMIC1 和其同源物新孢子虫 MIC1(NcMIC1)均优先结合α2-3-而非α2-6-连接的唾液酰-N-乙酰乳糖胺序列。然而,这三种凝集素在结合偏好上也存在差异。在胚胎细胞上报道的 TgMIC13 对α2-9 连接的二唾液酸序列的强烈结合以及在肠上皮上发现的相对强结合的 4-O-乙酰化-Sia 和 NcMIC1 对 6'-磺酰基唾液酰 Lewis(x)的结合可能对组织嗜性有影响。