蛋白芯片筛选鼠疫耶尔森菌与巨噬细胞的蛋白相互作用  

Screening protein-protein interactions between Yersinia pestis and host macrophage using protein chip

作　　者：杜宗敏[1] 谭亚芳[1] 李蓓[1] 童宗中[1] 江凌晓[1] 王津[1] 王红霞[1] 宋亚军[1] 郭兆彪[1] 杨瑞馥[1] 

机构地区：[1]军事医学科学院微生物流行病研究所,北京100071

出　　处：《军事医学科学院院刊》2005年第3期214-218,222,共6页Bulletin of the Academy of Military Medical Sciences

摘　　要：目的:利用蛋白芯片筛选鼠疫耶尔森菌的毒力因子,进一步揭示鼠疫菌与宿主之间相互作用的机制。方法:鼠疫菌毒力相关蛋白芯片筛选与小鼠巨噬细胞系774A.1可能有相互作用的蛋白质,免疫荧光试验验证筛选出的蛋白相互作用。结果:共筛选到17个可能与巨噬细胞有相互作用的鼠疫菌蛋白,并通过免疫荧光试验初步验证了其中8个蛋白芯片筛选试验中信号最强的蛋白与巨噬细胞的相互作用。结论:蛋白芯片技术可用于筛选病原菌与宿主之间的相互作用,有助于深入研究病原与宿主之间相互作用的机制。本研究筛选出的鼠疫菌蛋白与巨噬细胞的相互作用尚需深入研究和验证,以便最终确定这些蛋白在鼠疫菌致病机制中所发挥的作用。Objective: We attempt to use protein chip to screen protein-protein interactions between Yersinia pestis and macrophages, and reveal further clues for the mechanism of pathogenesis of this pathogen.Methods: Protein chip containing 149 recombinant virulence-related proteins of Y. pestis was used to investigate the protein-protein interactions between the pathogen and murine macrophage cell line J774A.1. The proteins that interact with macrophage proteins were probably the potential virulence factors and immuofluorescence assay and laser cofocal microscopy were performed to confirm the interactions. Results: Seventeen proteins showing interaction with proteins from macrophage were pick out as the candidates that are probably the virulence factors. Eight proteins showing the most intensive signals in protein screening were choosed to analyzed by immunofluorescence assay and their capability to interact with macrophage were confirmed.Conclusion: The results showed that protein chip is a very useful and high throughput tool for studying the protein interactions between the host and the pathogen. Based on these findings, further investigation can be done to illustrate the observed protein interactions screened by this work, to identify special targets in eukaryotic cell, and to reveal the significance, if any, of those interactions in pathogenesis.

关 键 词：耶尔森菌 鼠疫 巨噬细胞 蛋白质类 荧光免疫测定 

分 类 号：R378[医药卫生—病原生物学]