Large-Scale Identification of Bacteria-Host Crosstalk by Affinity Chromatography: Capturing the Interactions of Streptococcus suis Proteins with Host Cells
Large-Scale Identification of Bacteria-Host Crosstalk by Affinity Chromatography: Capturing the Interactions of Streptococcus suis Proteins with Host Cells
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DOI：
10.1021/pr200758q
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发表时间：
2011-11-01
影响因子：
4.4
通讯作者：
Jin, Meilin
中科院分区：
文献类型：
--
作者：
Chen, Bo;Zhang, Anding;Jin, Meilin
细菌与其宿主之间的蛋白质-蛋白质相互作用负责所有类型的感染过程。对细菌与宿主相互作用的研究有助于全面了解细菌性疾病的发病机制。尽管在这一领域的分散的努力，宿主和细菌蛋白质之间的相互作用的系统识别仍然不可用。在这里，我们开发ACSP（亲和色谱为基础的表面蛋白质组学），它结合亲和色谱和鸟枪蛋白质组学（LC-MS/MS），研究大规模的相互作用。应用ACSP技术，将Hep-2细胞表面分子固定化的层析树脂上，捕获猪链球菌2型（SS 2）的潜在表面相互作用蛋白（SIPs）。通过LC-MS/MS共鉴定出40个潜在的SIPs，其中3个为文献报道的SIPs。我们选择了8个重要的SIPs，并证实了它们粘附Hep-2细胞的能力。另外，发现3种新鉴定的SIPs或其多克隆抗体显著抑制SS 2与Hep-2细胞的粘附，表明它们在SS 2与Hep-2细胞之间的相互作用中起重要作用。使用这个例子，我们表明，ACSP是一个新的有价值的工具，调查细菌宿主相互作用。
Protein-protein interactions between bacteria and their hosts are responsible for all types of infection processes. The investigation of the bacteria host crosstalk can provide a comprehensive understanding of the pathogenesis of bacterial disease. Despite scattered efforts in this field, a systematic identification of interactions between host and bacterial proteins remains unavailable. Here, we develop ACSP (affinity chromatography-based surface proteomics), which combines affinity chromatography and shotgun proteomics (LC-MS/MS), to investigate the interactions on a large-scale. Using ACSP, the potential surface interacting proteins (SIPs) of Streptococcus suis serotype 2 (SS2) were captured by the chromatographic resin, which was immobilized with the native surface molecules of Hep-2 cells. And then 40, potential SIPs were identified from the preys by LC-MS/MS, including 3 SIPs that have been previously reported in the literature. We selected 8 important SIPs and confirmed their ability to adhere to Hep-2 cells. Additionally, 3 newly identified SIPs, or their polyclonal antibodies, were found to significantly inhibit the adherence of SS2 to Hep-2 cells, indicating their essential role in the interaction between SS2 and Hep-2 cells. Using this example, we show that ACSP represents a new valuable tool for investigating the bacteria host interactions.