利用FRET技术研究Hepcidin 和Fpn 相互作用

铁是生命必需的微量元素,ferroportin(Fpn)是小肠吸收细胞铁释放的重要蛋白。新近发现肝脏分泌的抗菌多肽 hepcidin 具有调节肠铁吸收的重要作用,但目前尚缺少Fpn和hepcidin发生作用的实验依据。应用荧光共振能量转移技术(fluorescence resonance energy transfer ,FRET)对hepcidin和Fpn之间的作用关系进行了深入研究。首先进行了hepcidin-CF P融合蛋白表达载体的构建及表达鉴定;然后对含YFP,Fpn-YFP基因动物细胞表达载体的构建、表达和FRET检测。实验结果证实hepcidin和Fpn之间存在直接的相互作用,并发现两种蛋白发生相互作用后hepcidin也在细胞质中有分布。为临床治疗铁代谢紊乱性疾病提供了新的治疗策略和重要理论依据。

The Study of the Interaction Between Hepcidin and Fpn by FRET

Iron is an essential trace element of life, ferroportin (Fpn) is the intestinal absorption of iron release of important cell proteins. Newly discovered antimicrobial peptide hepcidin secreted by the liver can regulate the important role of intestinal iron absorption, but still play a role in the lack of Fpn and experimental basis for hepcidin. So fluorescence resonance energy transfer technology (fluorescence resonance energy transfer, FRET) on the role of hepcidin and Fpn relationship between the depth study. First of all, were hepcidin-CFP fusion protein expression vector and identification; then with YFP, Fpn-YFP gene in animal cell expression vector, expression, and FRET detection. The results confirmed that hepcidin and Fpn direct interaction between, and found that two proteins interact in the cytoplasm after the distribution of hepcidin also. The results for the treatment of disorders of iron metabolism to provide a new and important theoretical basis for therapeutic strategies.