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TRAF2是机体免疫与炎症反应中具有重要作用的蛋白,有E3连接酶活性;GFP是一种受到激发光照射后可产生绿色荧光的蛋白,常用于蛋白质的细胞共定位研究,并显示与GFP融合表达靶蛋白在细胞中的位置.我们实验发现共转染表达质粒pCMV-myc-TRAF2与pEGFP-C3,可引起转染细胞绿色荧光减弱.Western印迹实验证明TRAF2可以降解GFP,并且这种降解是通过蛋白酶体途径进行的.为进一步确定这种降解的特异性,在HEK293细胞中共转pCMV-myc-TRAF2与pEGFP-C3-LNX后,发现随着TRAF2表达量增加,融合蛋白GFP-LNX减少;而共转质粒pCMV-myc-TRAF2与pCMV-myc-LNX后,未发现LNX蛋白表达减少,表明TRAF2对GFP的降解具有相对特异性.GFP是人类细胞中并不存在的蛋白,TRAF2能够将其降解可能意味着TRAF2参与了细胞抗病原体感染的过程.
TRAF2 is a protein that plays an important role in the immune and inflammatory response of the body and has E3 ligase activity. GFP is a green fluorescent protein that is irradiated by excitation light and is commonly used in cell colocalization of proteins and has been shown to be fused with GFP Expression of the target protein in the cell location.Our experiments found that co-transfected expression plasmid pCMV-myc-TRAF2 and pEGFP-C3, can cause transfected cells decreased green fluorescence.Western blotting experiments show that TRAF2 can degrade GFP, and this degradation is To further confirm the specificity of this degradation, after co-transfecting pCMV-myc-TRAF2 and pEGFP-C3-LNX in HEK293 cells, it was found that the fusion protein GFP-LNX decreased as the expression of TRAF2 increased; However, no expression of LNX protein was found after pCMV-myc-TRAF2 and pCMV-myc-LNX were co-transfection, indicating that TRAF2 is relatively specific to GFP degradation.GFP is a non-existing protein in human cells, Degradation may mean that TRAF2 is involved in the cellular anti-pathogen infection process.