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Amer1(APC membrane recruitment protein 1)又称为WTX(Wilms’tumor X),是首个发现位于X染色体上的抑癌基因.由于其定位的特殊性,Amer1近年来成为研究的热点之一.研究表明,Amer1作为骨架蛋白在细胞内与多种蛋白(APC,β-catenin,Axin等)直接结合,在Wnt信号通路中发挥着重要功能.据报道Amer1/WTX含有3个肿瘤抑制蛋白质APC(adenomatous polyposis coli,APC)的结合位点,对于APC在细胞膜上的定位过程中发挥着重要的作用.但是,通过序列比对发现,Amer1可能存在第4个被忽略的APC的结合位点,定位于A1和A2之间.为了验证该片段能否与APC结合,分别构建了GST-Amer1(365-375)和His-APC(407-775)两种重组蛋白.通过GSTpull down,证明了这两个片段存在相互作用,并进一步通过ITC(isothermal titration calorimetry)实验测定了两者结合的亲和力.本研究结果不仅在体外证实了Amer1第4个APC结合位点的存在,也为APC和Amer1/WTX复合物的结构和功能的研究打下了良好的基础.
Amer1 (APC membrane recruitment protein 1), also known as WTX (Wilms’tumor X), is the first tumor suppressor gene found on chromosome X. Due to the specificity of its location, Amer1 has become one of the hot topics in recent years. These results suggest that Amer1, as a scaffold protein, binds directly to various proteins (APC, β-catenin, Axin, etc.) and plays an important role in the Wnt signaling pathway.Amer1 / WTX is reported to contain three tumor suppressor proteins, APC polyposis coli, APC) plays an important role in the localization of APC on the cell membrane.However, through sequence alignment, Amer1 may have the 4th neglected APC binding site located in Between A1 and A2, two recombinant proteins, GST-Amer1 (365-375) and His-APC (407-775), were constructed in order to verify whether this fragment can bind to APC.According to GSTpull down, And their binding affinities were further determined by an isothermal titration calorimetry (ITC) assay.The results of this study not only confirmed the presence of the fourth APC binding site of Amer1 in vitro but also the combination of APC and Amer1 / WTX The structure and function of things Able to lay a good foundation for research.