Interaction of Prevotella intermedia strain 17 leucine-rich repeat domain protein AdpF with eukaryotic cells promotes bacterial internalization.
Interaction of Prevotella intermedia strain 17 leucine-rich repeat domain protein AdpF with eukaryotic cells promotes bacterial internalization.
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中间普雷沃氏菌菌株 17 富含亮氨酸的重复结构域蛋白 AdpF 与真核细胞的相互作用促进细菌内化。
DOI：
10.1128/iai.01361-13
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发表时间：
2014
影响因子：
3.1
通讯作者：
Lewis,JaninaP
中科院分区：
文献类型：
--
作者：
Sengupta,Dipanwita;Kang,Dae-Joong;Anaya-Bergman,Cecilia;Wyant,Tiana;Ghosh,ArnabK;Miyazaki,Hiroshi;Lewis,JaninaP
中间普雷沃氏菌是一种与多种口腔疾病有关的口腔细菌。虽然这种细菌的非吞噬宿主细胞的内化是很好地建立，介导的过程中的分子球员不为人所知。在这里，一个富含亮氨酸的重复序列（LRR）结构域蛋白，指定AdpF的属性，进行了描述。该蛋白含有由663个氨基酸残基组成的富含亮氨酸的区域，分子模拟显示其折叠成经典的弯曲螺线管结构。通过电子显微镜和共聚焦显微镜分析证实了重组AdpF（rAdpF）的细胞表面定位。该蛋白的重组形式以剂量依赖性方式结合纤连蛋白。此外，该蛋白质被内化的宿主细胞，在30分钟内完成的大部分过程。rAdpF的内化被抑制制霉菌素，细胞松弛素，latrunculin，诺考达唑，渥曼青霉素，表明微管，微丝，和信号转导所需的入侵。值得注意的是，真核细胞与AdpF的预孵育分别使HeLa和NIH 3T3成纤维细胞系的中间毕赤酵母17内化增加了5倍和10倍。rAdpF蛋白的加入也非常有效地诱导细菌内化到口腔上皮细胞系HN 4中，以及内化到原代细胞中，包括人口腔角质形成细胞（HOK）和人脐静脉内皮细胞（HUVEC）。最后，暴露于中间青霉17的细胞在再感染时更容易内化细菌。综上所述，我们的数据表明，rAdpF在各种宿主细胞内化中间毕赤酵母17中起作用。
Prevotella intermedia is an oral bacterium implicated in a variety of oral diseases. Although internalization of this bacterium by nonphagocytic host cells is well established, the molecular players mediating the process are not well known. Here, the properties of a leucine-rich repeat (LRR) domain protein, designated AdpF, are described. This protein contains a leucine-rich region composed of 663 amino acid residues, and molecular modeling shows that it folds into a classical curved solenoid structure. The cell surface localization of recombinant AdpF (rAdpF) was confirmed by electron and confocal microscopy analyses. The recombinant form of this protein bound fibronectin in a dose-dependent manner. Furthermore, the protein was internalized by host cells, with the majority of the process accomplished within 30 min. The internalization of rAdpF was inhibited by nystatin, cytochalasin, latrunculin, nocodazole, and wortmannin, indicating that microtubules, microfilaments, and signal transduction are required for the invasion. It is noteworthy that preincubation of eukaryotic cells with AdpF increased P. intermedia 17 internalization by 5- and 10-fold for HeLa and NIH 3T3 fibroblast cell lines, respectively. The addition of the rAdpF protein was also very effective in inducing bacterial internalization into the oral epithelial cell line HN4, as well as into primary cells, including human oral keratinocytes (HOKs) and human umbilical vein endothelial cells (HUVECs). Finally, cells exposed to P. intermedia 17 internalized the bacteria more readily upon reinfection. Taken together, our data demonstrate that rAdpF plays a role in the internalization of P. intermedia 17 by a variety of host cells.