中国药理学通报2016，Vol.32Issue(12)：1761-1766,6.DOI:10.3969/j.issn.1001-1978.2016.12.024
大肠不耐热肠毒素B亚单位佐剂活性相关蛋白的筛选
Exploration for interacting protein of E.coli heat-labile enterotoxin B subunit(LTB)as adjuvant
摘要
Abstract
Aim To explore the mechanism of E.coli heat-labile enterotoxin B subunit(LTB)as adjuvant by analysis of cellular proteins interacting with LTB. Methods Whole cell proteins were purified from RAW 264.7 cell after treated with LTB or NaCl 12 h, respectively.The cellular proteins were interacted with LTB and the interacting proteins were purified by pull-down assay and identified by mass spectrography.The LTB interaction proteins were conformed with Western blot and immunofluorescence assay.Results 25 LTB interaction proteins were found,and their interaction network was mapped;four proteins (Jup,Dsp,Ddx5 and Vimentin)were indicated to be related with LTB adjuvant activity;immunofluorescence assay indicated that GM130 interacted with LTB,however,Vimentin had no interaction with LTB in vivo.After treated by LTB,the expression of β-actin was upregulated obvi-ously in RAW 264.7 cell,whereras,Hspd1 did not show any change.Conclusions LTB exerts adjuvant activity through binding to GM1 of immune cells,cau-sing endocytosis and transporting to the Golgi apparatus by vesicles.Then LTB might bind to Jup and affect TCF/LEF activity,regulating the expression of Bcl 2, IL-6,and Runx3.The result is promoted T cell and B cell proliferation,differentiation and activation by se-cretion of cytokines and immunoglobulins.关键词
LTB/RAW264.7/相互作用蛋白/免疫/佐剂/内吞Key words
LTB/RAW 264.7/interacting protein/immune/adjuvant/endocytosis引用本文复制引用
刘林,周慧聪,王秋娟,陈思静,马永平..大肠不耐热肠毒素B亚单位佐剂活性相关蛋白的筛选[J].中国药理学通报,2016,32(12):1761-1766,6.基金项目
国家自然科学基金资助项目 （）
