沙眼衣原体包涵体膜蛋白CT225与宿主波形蛋白存在相互作用

【背景】衣原体独特的发育周期是在包涵体内完成的,大约7%–10%的基因编码包涵体膜蛋白,由此可见包涵体膜蛋白可能在其发育和致病过程中发挥重要作用。然而,其具体功能仍有待深入研究。【目的】筛选包涵体膜蛋白CT225的互作分子,以期进一步了解其可能的生物学功能。【方法】首先表达融合蛋白GST-CT225,用亲和层析法从HeLa细胞裂解液中筛选CT225的互作分子,所得蛋白进行质谱分析。确定候选蛋白,然后通过免疫共沉淀方法(Co-Immunoprecipitation,CO-IP)、谷胱甘肽巯基转移酶(Glutathione S-Transferase,GST)下拉/沉降实验和亚细胞定位等方法进行验证。【结果】参考质谱分析得分,通过实验初步验证得出波形蛋白(Vimentin,VIM)为与CT225相互作用的蛋白。【结论】CT225与HeLa细胞的波形蛋白Vimentin互相作用,提示其功能可能与维持细胞骨架完整性、膜运输和脂质转运等有关。

Chlamydia trachomatis CT225 interacting with vimentin from HeLa cell

[Background]The unique development cycle of Chlamydia trachomatis is completed in inclusion.About 7%-10%genes encode membrane proteins in the inclusion.Therefore,the membrane proteins in the inclusion may play an important role in the development and pathogenesis of Chlamydia.However,its specific functions still need to be further studied.[Objective]In order to further understand the function of the membrane protein CT225 in the inclusion,the association molecules were screen.[Methods]The fusion protein GST-CT225 was first expressed.Then,the interaction molecules of CT225 were screened from the lysate of HeLa cell by affinity chromatography,and analyzed by mass spectrometry.Finally,the candidate proteins were selected and verified by Co-immunoprecipitation,glutathione S-transferase(GST)pull-down test and subcellular localization.[Results]Vimentin was the protein interacted with CT225 according to the score of mass spectrometry.[Conclusion]CT225 interacts with vimentin in HeLa cells,suggesting that CT225 has the relationship with the cytoskeleton integrity,membrane transport and lipid transport.