精子膜蛋白YWK-Ⅱ与穆勒管抑制物质的结合及结合区域的确定

Binding of Human Sperm Transmembrane Protein YWK-Ⅱwith Müllerian Inhibiting Substance and Determination of the Binding Site

目的验证精子膜蛋白YWK-Ⅱ与穆勒管抑制物质（MIS）的结合作用, 并确定在MIS上的结合区域。方法将YWK-Ⅱ蛋白胞外区一段63个氨基酸的片段在大肠杆菌中表达并纯化，运用Pull-down实验研究纯化的蛋白质与MIS的作用，并采用酵母双杂交系统确定YWK-Ⅱ蛋白在MIS上的结合区域。结果重组YWK-Ⅱ蛋白能与大鼠睾丸提取液中的MIS结合，结合部位位于MIS 的N肽段中一段较保守区域内。结论精子膜上的YWK-Ⅱ蛋白可能是卵巢分泌的MIS的受体。

Objective To verify the interaction between the YWK-Ⅱprotein and Müllerian inhibiting substance (MIS) and determine the binding site on MIS. Methods A 63 amino acids segment of the extracellular domain of YWK-Ⅱprotein was expressed in E. coli and was purified. The interaction between the purified protein and MIS was analyzed by Pull-down assay. Yeast two-hybrid system was used to determine the binding site on MIS. Results The recombinant YWK-Ⅱprotein could interact with MIS in rat testis extracts. The binding site was located at a conserved segment of MIS. Conclusion The YWK-Ⅱprotein might act as the receptor of MIS secreted by ovary.