Sperm proteases that may be involved in the initiation of sperm motility in the newt, Cynops pyrrhogaster.
Sperm proteases that may be involved in the initiation of sperm motility in the newt, Cynops pyrrhogaster.
复制标题
DOI：
10.3390/ijms150915210
复制
发表时间：
2014-08-28
影响因子：
5.6
通讯作者：
Watanabe A
中科院分区：
文献类型：
--
作者：
Yokoe M;Sano M;Shibata H;Shibata D;Takayama-Watanabe E;Inaba K;Watanabe A
蝾螈Cynopspyrrhogaster精子在顶体反应（AR）后释放的蛋白酶被认为能够裂解卵胶外表面的片状结构并释放精子运动启动物质（SMIS）。在这里，我们发现精子头部的蛋白酶活性能够有效地广泛消化精子下方的底物。在 AR 后的精子上清液中检测到通过荧光素硫代氨基甲酰-酪蛋白消化测量的蛋白酶活性，并且该活性被丝氨酸或半胱氨酸蛋白酶抑制剂 4-(2-氨基乙基)苯磺酰氟 (AEBSF) 抑制，表明 AR 释放了丝氨酸和/或半胱氨酸蛋白酶。在对睾丸的计算机分析中，顶体蛋白和 20S 蛋白酶体被确定为顶体蛋白酶的可能候选者。我们还在精子表面检测到另一种 AEBSF 敏感的蛋白酶活性。用 AlexaFluor 488 标记的 AEBSF 进行荧光染色，显示主片段中有半胱氨酸蛋白酶；它位于轴杆和波动膜之间的连接区域，其中包括轴丝并产生强大的膜波动以实现精子的向前运动。这些结果表明，顶体和主片中的 AEBSF 敏感蛋白酶可能参与卵果冻表面精子运动的启动。
A protease of sperm in the newt Cynops pyrrhogaster that is released after the acrosome reaction (AR) is proposed to lyse the sheet structure on the outer surface of egg jelly and release sperm motility-initiating substance (SMIS). Here, we found that protease activity in the sperm head was potent to widely digest substrates beneath the sperm. The protease activity measured by fluorescein thiocarbamoyl-casein digestion was detected in the supernatant of the sperm after the AR and the activity was inhibited by 4-(2-aminoethyl) benzenesulfonyl fluoride (AEBSF), an inhibitor for serine or cysteine protease, suggesting the release of serine and/or cysteine proteases by AR. In an in silico analysis of the testes, acrosins and 20S proteasome were identified as possible candidates of the acrosomal proteases. We also detected another AEBSF-sensitive protease activity on the sperm surface. Fluorescence staining with AlexaFluor 488-labeled AEBSF revealed a cysteine protease in the principal piece; it is localized in the joint region between the axial rod and undulating membrane, which includes an axoneme and produces powerful undulation of the membrane for forward sperm motility. These results indicate that AEBSF-sensitive proteases in the acrosome and principal piece may participate in the initiation of sperm motility on the surface of egg jelly.
登录
查看更多内容
影响因子：
0.9
作者：
Ukita, M;Itoh, T;Onitake, K
通讯作者：
Onitake, K
DOI：
10.1530/jrf.0.0210501
发表时间：
1970-01-01
期刊：
JOURNAL OF REPRODUCTION AND FERTILITY
影响因子：
--
作者：
ALLISON, AC;HARTREE, EF
通讯作者：
HARTREE, EF
影响因子：
3.6
作者：
MACK, S;BHATTACHARYYA, AK;ZANEVELD, LJD
通讯作者：
ZANEVELD, LJD
DOI：
10.1093/protein/7.1.57
发表时间：
1994-01-01
期刊：
PROTEIN ENGINEERING
影响因子：
--
作者：
RYPNIEWSKI, WR;PERRAKIS, A;WILSON, KS
通讯作者：
WILSON, KS
影响因子：
3.8
作者：
Sasanami, Tomohiro;Sugiura, Kenichi;Matsuda, Tsukasa
通讯作者：
Matsuda, Tsukasa