Siegel M S, Bechtold D S, Kopta C I, Polakoski K L
Biol Reprod. 1986 Sep;35(2):485-91. doi: 10.1095/biolreprod35.2.485.
The proacrosin-acrosin proteinase system was measured and partially characterized in unpurified extracts of washed hamster epididymal sperm. Autoactivation experiments demonstrated that proacrosin accounted for greater than 98% of the acrosin activity in the sperm extracts from individual animals. Several bands of proteinase activity were observed on gelatin-sodium dodecyl sulfate-polyacrylamide gel electrophoretic (gelatin-SDS-PAGE) zymography. The major proteinase activities in the nonactivated extracts corresponded to relative molecular masses (Mr) of 51,000 to 56,000, while less distinct digestion occurred with relative molecular masses of 37,000 to 49,000. It was demonstrated that after a serial dilution of the sperm extract, the proteinase activity in as few as 6,000 sperm could readily be detected by the gelatin-SDS-PAGE methods. Time-course activation studies showed that the zymogen was completely converted to active proteinase in 45-60 min at pH 8.0 and 25 degrees C. This autoconversion process was markedly inhibited by calcium, sodium, and heparin. However, each of these compounds stimulated the proteolytic activity of acrosin. These studies demonstrate that the proacrosin-acrosin system can be investigated in extracts of nonpurified hamster epididymal sperm.
在洗涤过的仓鼠附睾精子的未纯化提取物中对前顶体蛋白酶 - 顶体蛋白酶系统进行了测定并部分表征。自激活实验表明，前顶体蛋白酶在单个动物的精子提取物中占顶体蛋白酶活性的98%以上。在明胶 - 十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳（明胶 - SDS - PAGE）酶谱上观察到几条蛋白酶活性带。未激活提取物中的主要蛋白酶活性对应于相对分子质量（Mr）为51,000至56,000，而相对分子质量为37,000至49,000时消化不太明显。结果表明，在对精子提取物进行系列稀释后，通过明胶 - SDS - PAGE方法可以很容易地检测到低至6000个精子中的蛋白酶活性。时间进程激活研究表明，在pH 8.0和25℃下，酶原在45 - 60分钟内完全转化为活性蛋白酶。这种自动转化过程受到钙、钠和肝素的显著抑制。然而，这些化合物中的每一种都刺激了顶体蛋白酶的蛋白水解活性。这些研究表明，可以在未纯化的仓鼠附睾精子提取物中研究前顶体蛋白酶 - 顶体蛋白酶系统。
