摘要
我们运用 Percoll密度梯度离心技术从正常捐精者的精液中分离出成熟精子并使其获能 ,经含 1 %穿通 (Triton) X 1 1 4Tris缓冲液溶解 ,超声粉碎及离心分离后获得人精子可溶性蛋白悬液。随后运用蛋白质亲和层析及 SDS PAGE电泳技术分离人精子可溶性蛋白质 ,经银染后发现 :用山羊抗人巨噬细胞甘露糖受体 (anti macrophage mannosereceptor,AMMR) Ig G包被的 Affi Prep Hz support珠能结合人精子中一种分子量为2 9KD的蛋白质 ,而对照组为正常山羊血清 (normal goat,NG) Ig G包被珠 ,其结果却呈阴性。最后运用精子膜生物素标记及蔗糖密度梯度离心技术进一步标记分离人精子膜可溶性蛋白质 ,证实了这种与 AMMR Ig G结合的蛋白质为甘露糖结合蛋白 ,存在于人精子膜上。精子膜甘露糖结合蛋白可能与精卵识别的起始过程有相当密切的关系。
Using affinity chromatography and sodium dodecyl sulphatepolyacrylamide gel electrophoresis (SDSPAGE) methods, we found that the beads coupled with goat antihuman macrophage mannose receptor IgG could bind one kind of human spermatozoan proteins whose molecular weight was about 29 KD,while the beads coupled with normal goat IgG could not. Employing the centrifugal technique of sucrose density gradient and the technique of spermatozoan membrane biotinylation to separate the spermatozoan membrane proteins, we also verified that the protein which had affinity to the beads coupled with goat antimacrophage mannose receptor IgG belonged to spermatozoan membrane proteins. The mannose binding protein on the spermatozoan membrane may play an important role in spermegg recognition during fertilization.
出处
《生殖与避孕》
CAS
CSCD
北大核心
2000年第1期20-25,共6页
Reproduction and Contraception
关键词
精子膜
甘露糖结合蛋白
亲和层析
Spermatozoan membrane, Mannose binding protein, Chromatography
