新疆农业科学2017，Vol.54Issue(7)：1284-1290,7.DOI:10.6048/j.issn.1001-4330.2017.07.014
卵泡刺激素功能肽与受体胞外区不同片段结合作用的分析
Analysis of the Binding Effect of Follicle Stimulating Hormone Peptide and Different Fragments of Extracellular Domain
摘要
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.关键词
FSH33-53肽/LRR/FSHR9-30肽/胞外区/亲和力Key words
FSH33-53 peptides/LRR/FSHR9-30 peptides/ECD,affinity分类
农业科技引用本文复制引用
赵婷,席欧彦,秦瑞坪,邱玲玲,马晓玲,李江伟..卵泡刺激素功能肽与受体胞外区不同片段结合作用的分析[J].新疆农业科学,2017,54(7):1284-1290,7.基金项目
国家自然科学基金项目"抗卵泡刺激素受体纳米抗体的制备及其在肿瘤分子显像和抗血管治疗中的作用"(81260333) National Natural Science Foundation of China "Preparation of Nano Antibody against Follicle Stimulating Hormone Receptor and Its Role in Tumor Molecular Imaging and Angiogenesis Therapy(81260333) （81260333）
