兔子宫β-糖蛋白的进一步特性研究。

Further characterization of the beta-glycoprotein of the rabbit uterus.

作者信息

Thie M, Bochskanl R, Kirchner C

出版信息

Biol Reprod. 1986 Oct;35(3):683-9. doi: 10.1095/biolreprod35.3.683.

DOI:10.1095/biolreprod35.3.683

Abstract

beta-Glycoprotein was isolated from preimplantation uterine secretions of the rabbit by gel- and ion-exchange chromatography. Two fractions, called DF1 and DF2, were analyzed by isoelectric focusing (IEF) and sodium-dodecyl-sulfate polyacrylamide gel electrophoresis (SDS-PAGE) in combination with Western blotting and immunoelectrophoresis. DF1 displayed 21 bands with isoelectric points of pH 5.2-7.6, and DF2 15 bands of pH 4.2-5.7. SDS-PAGE yielded up to 14 bands with major components at molecular weights of 63,000 and 135,000 respectively. Two-dimensional gel electrophoresis of DF2 in combination with Western blotting revealed five groups of proteins of equal molecular weights but with different isoelectric points, indicating immunological identities. Glycosidase activities in uterine secretions before and after implantation were studied and compared with those of the blastocyst fluids. alpha-L-Fucosidase co-eluted with DF1, and beta-N-acetylglycosaminidase was distributed in DF1 and DF2. Both enzymes were localized on isoelectric focusing gels, and N-acetylglucosaminidase was also demonstrated in an immunoprecipitate of DF1.

摘要

通过凝胶和离子交换色谱法从兔植入前子宫分泌物中分离出β-糖蛋白。对称为DF1和DF2的两个组分进行等电聚焦（IEF）、十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE）分析，并结合蛋白质印迹法和免疫电泳法。DF1显示出21条等电点为pH 5.2 - 7.6的条带，DF2显示出15条pH 4.2 - 5.7的条带。SDS-PAGE产生多达14条带，主要成分的分子量分别为63,000和135,000。DF2的二维凝胶电泳结合蛋白质印迹法显示出五组分子量相等但等电点不同的蛋白质，表明它们具有免疫同一性。研究并比较了植入前后子宫分泌物中的糖苷酶活性以及与囊胚液中的糖苷酶活性。α-L-岩藻糖苷酶与DF1共洗脱，β-N-乙酰氨基葡糖苷酶分布于DF1和DF2中。两种酶都定位在等电聚焦凝胶上，并且在DF1的免疫沉淀物中也证实了N-乙酰葡糖胺糖苷酶的存在。