NADE与MATH2蛋白相互作用的研究

Preliminary study on the interaction between human proteins NADE and MATH2

目的探讨NADE与MATH-2蛋白之间相互作用及MATH-2对NADE蛋白功能的调控效应。方法用GST-pulldown与免疫共沉淀(CoIP)和细胞共定位方法验证NADE与MATH-2蛋白之间的相互作用,用通过流式细胞仪检测蛋白相互作用对凋亡的影响。结果NADE与MRTH2蛋白存在相互作用,这两个蛋白能共定位于SH-SY5Y细胞的胞浆中。二者之间的相互作用能够降低UV引起的细胞凋亡。结论研究首次证实了NADE与MATH2之间的相互作用,发现该相互作用可能通过干扰NADE对紫外线诱发的细胞凋亡产生影响。

Objective The interaction between human proteins NADE and MATH2 was examined. Methods The human NADE was used as a bait to search for partners in human brain eDNA library by yeast two-hybrid screening. The interaction was confirmed in vitro and in vivo by GST pull-down and co-immuno- precipitation, and subeellular co-localization was carried out in SH-SY5Y cells. Fluorescence activated cell sorting （FCAS） was used to examine the influence of this interaction on UV-induced apoptosis in 293T cells. Results The protein interaction was validated both in vitro and in vivo. Moreover, the subcellular localization assay showed that the two proteins could co-localize in the cytoplasm of SH-SY5Y cells. Co-transfeetion of MATH2 and NADE protects against UV-induced apoptosis in 293T cells. Conclusions A new protein interaction between NADE and MATH2 was identified. The interaction exerts impact on UV-induced apoptosis in 293T cells.