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人精浆中与精子前向运动相关蛋白的研究——蛋白的分离及活性检测

Identification of sperm forward motility related proteins from human seminal plasma-protein isolation and characterization

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【作者】 丁之德高惠宝吴明章张永莲

【Author】 Ding Zhide 1,Gao Huibao 1,Wu Mingzhang 1,Zhang Yonglian 2 1Dept.of Histology and Embryology,Shanghai Second Medical University,(Shanghai 200025); 2 Shanghai Institute of Biochemical,Chinese Academy of Science

【机构】 上海第二医科大学组织胚胎学教研室!上海200025中科院上海生物化学研究所

【摘要】 我们运用ConA包被珠的蛋白质亲和层析分离技术、蛋白质超滤技术对热处理后的人精浆进行蛋白分离、浓缩 ,结果发现 :蛋白亲和层析洗脱浓缩液加茶碱的实验组与公牛附睾头部的精子共同孵育后 ,经计算机辅助的精子分析技术显示精子的前向运动速度显著高于仅用茶碱的对照组 (P <0 .0 1)。蛋白亲和层析洗脱浓缩液经聚丙酰胺凝胶电泳和SDS 聚丙酰胺凝胶电泳及Coomassieblue染色后显示 :蛋白条带由聚丙酰胺凝胶中的高分子量解离成SDS 聚丙酰胺凝胶中的低分子量 ,经SDS 聚丙酰胺凝胶电泳分离的主要蛋白其分子量分别为 46 .8和 5 7.5kDa。精浆中与精子运动相关蛋白的分离及深入的研究对进一步了解其作用机制具有十分重要的基础理论和临床意义

【Abstract】 Employing techniques of the affinity chromatography covered with ConA beads and protein ultrafiltration,some proteins were isolated and concentrated from the heated human seminal plasma.The results of computer assistant semen analyses (CASA)revealed that the forward motility index of spermatozoa from epididymal caput of bovine incubated with proteins and theophylline was significantly different with that of spermatozoa incubated with theophylline only ( P <0.01).The index of former was obviously higher than that of latter.Using the methods of PAGE,SDS?PAGE and Coomassie blue staining,the protein bands with high molecules in the gel of PAGE changed into low molecules in the gel of SDS?PAGE.The major bands of the proteins in the gel of SDS?PAGE were 46.8 and 57.5 kDa.The proteins related to the forward motility of spermatozoa in mammalian seminal plasma play some vital roles during the maturation of spermatozoa through the epididymis and fertilization in the female reproductive tract.

  • 【文献出处】 中国男科学杂志 ,Chinese Journal of andrology , 编辑部邮箱 ,2001年02期
  • 【分类号】R321.1
  • 【被引频次】3
  • 【下载频次】119
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