白血病相关蛋白16与角蛋白18相互作用的鉴定

目的：验证白血病相关蛋白（LRP）16与角蛋白（KRT）家族成员KRT18之间的相互作用关系。方法：PCR扩增KRT18及其结构域缺失体基因片段,构建KRT18全长及其结构域真核表达载体,通过体外GST pull down实验和体内免疫共沉淀实验验证LRP16/KRT18的相互作用,并鉴定其作用的结构域。结果：构建了KRT18及其结构域重组子,GST pull down实验证明LRP16与KRT18在体外存在相互作用,它们的特异结合区域位于KRT18的C端,KRT18能够与LRP16的C端结构域相互作用;免疫共沉淀实验表明内源性LRP16与KRT18的C端在体内存在特异结合。结论：KRT18与LRP16存在相互作用,为进一步研究KRT18影响LRP16的亚细胞分布及调控LRP16核功能执行的作用机制奠定了基础。

Interaction Between Leukemia Related Protein 16 and Keratin 18

Objective： To further confirm the interaction of leukemia related protein（LRP） 16 with keratin（KRT） 18. Methods： Constructing recombinant expression vector of KRT18 full length and its function structural domains, and the interaction between LRP16 and KRT18 was confirmed by GST pull-down assay in vitro and co-immunoprecipitation analysis in vivo. In addition, the interacting domains of both LRP16 and KRT18 were checked by GST pull-down Results： The recombinant expression vector of KRT18 and its function struction domains were constructed. LRP16 has direct interactions with KRT18. The unique domain in C terminal of LRP16 interacts with C terminal region of KRT18 protein. Conclusion： These results propose reliable experimenta data for the molecular interaction of LRP16 with KRT18 in vitro and in vivo, and provide foundation for further study of functional regulation of LRP16.