Abstract: [Objective] The FSH33-53 peptide binds to its receptor FSHR and activates the downstream signal as a FSH functional peptide.However, its specific binding position on the receptor is unclear.The aim of this study is to elucidate the binding region of the FSH33-53 peptide on the receptor FSHR in the hope of providing a basis for FSH-based vaccine design.[Method]The extracellular domain(ECD)and leucine-rich repeat (LRR) of follicle-stimulating hormone receptor was amplified by polymerase chain reaction (PCR) and constructed recombinant plasmids pET22b-FSHR-ECD and pET22b-FSHR-LRR.Protein FSHR-ECD and FSHR-LRR were obtained by expression and purification.FSHR9-30-KLH.was obtained by peptide synthesis, and binding and affinity of receptor fragments with FSH33-53 peptide were detected by ELISA method.[Result]The proteins FSHR-ECD and FSHR-LRR were successfully expressed and purified and their relative molecular mass (MR) was 43 and 32kda., respectively.When the receptor was 0.5 μg / mL and the ligand was 2.5 μg / mL, the three proteins were bound to the FSH33-53 peptide.The affinity of ELISA to detect the ligand and ligand was 0.21 × 10-6, 0.45 × 10-6 and 0.056 × 10-6 mol / L, respectively.[Conclusion]The binding of LRR fragment to FSH33-53 peptide is stronger than that of the other two fragments.

摘要： [目的]卵泡刺激素(FSH)33-53肽可以与FSH受体(FSHR)结合,并激活下游信号,被作为FSH功能肽.然而其在受体上的具体结合位置还不清楚.阐明FSH33-53肽在FSHR上的结合区域,为基于FSH的疫苗设计提供依据.[方法]采用重组的FSHR 胞外区(ECD)和富含亮氨酸区(LRR)及合成的FSHR9-30区分析这些FSHR的功能区域与FSH33-53的结合作用.PCR扩增得到hFSHR ECD和hFSHR LRR目的基因,构建重组质粒pET22b-FSHR-ECD和pET22b-FSHR-LRR,表达纯化获得蛋白FSHR-ECD和FSHR-LRR.采用多肽合成法获得FSHR9-30-KLH.通过ELISA方法检测受体片段与FSH33-53肽的结合和亲和力.[结果]获得FSHR-ECD和FSHR-LRR蛋白,其相对分子质量(Mr)分别为43和32kDa.在受体为0.5 μg/mL,配体为2.5 μg/mL时,三种蛋白均与FSH33-53肽结合.ELISA检测受体与配体的亲和力分别为0.21×10-6、0.45×10-6和0.056×10-6 mol/L .[结论]FSHR 9-30片段与FSH33-53肽的结合较其它两个片段结合力强.