Identification, characterization, and functional correlation of calmodulin-dependent protein phosphatase in sperm.
Identification, characterization, and functional correlation of calmodulin-dependent protein phosphatase in sperm.
复制标题
精子中钙调蛋白依赖性蛋白磷酸酶的鉴定，表征和功能相关性。
DOI：
10.1083/jcb.106.5.1625
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发表时间：
1988-05
期刊：
影响因子：
--
通讯作者：
Means AR
中科院分区：
文献类型：
--
作者：
Tash JS;Krinks M;Patel J;Means RL;Klee CB;Means AR
初步数据表明，钙对重新激活的精子活力的抑制与抑制蛋白磷酸化有关。钙调素依赖性蛋白磷酸酶（钙调磷酸酶）催化的磷酸化的抑制被发现。来自狗、猪和海胆的精子含有该酶的Ca 2+结合B亚基（Mr 15，000）和钙调蛋白结合A亚基（Mr 63，000）。精子A亚基是略高于先生比其他组织的报告。内源性钙调素依赖性蛋白磷酸酶活性与单特异性抗体的抑制揭示了14磷蛋白底物在精子中的这种酶的存在。该酶定位于鞭毛和精子头的顶体后区。鞭毛磷酸酶活性定量提取与0.6 M KCl从分离的鞭毛从狗，猪，和海胆精子。所有的盐可提取的磷酸酶活性抑制与抗体对正宗的酶。预孵育的精子模型与纯化的磷酸酶刺激的曲线速度和横向头振幅（超活化游泳模式的重要组成部分）和抑制节拍交叉频率表明这种酶在轴丝功能的作用。我们的研究结果表明，钙调素依赖性蛋白磷酸酶在鞭毛运动的钙依赖性调节中起着重要作用。
Preliminary data demonstrated that the inhibition of reactivated sperm motility by calcium was correlated with inhibited protein phosphorylation. The inhibition of phosphorylation by Ca2+ was found to be catalyzed by the calmodulin-dependent protein phosphatase (calcineurin). Sperm from dog, pig, and sea urchin contain both the Ca2+-binding B subunit of the enzyme (Mr 15,000) and the calmodulin- binding A subunit with an Mr of 63,000. The sperm A subunit is slightly higher in Mr than reported for other tissues. Inhibition of endogenous calmodulin-dependent protein phosphatase activity with a monospecific antibody revealed the presence of 14 phosphoprotein substrates in sperm for this enzyme. The enzyme was localized to both the flagellum and the postacrosomal region of the sperm head. The flagellar phosphatase activity was quantitatively extracted with 0.6 M KCl from isolated flagella from dog, pig, and sea urchin sperm. All salt-extractable phosphatase activity was inhibited with antibodies against the authentic enzyme. Preincubation of sperm models with the purified phosphatase stimulated curvolinear velocity and lateral head amplitude (important components of hyperactivated swimming patterns) and inhibited beat cross frequency suggesting a role for this enzyme in axonemal function. Our results suggest that calmodulin-dependent protein phosphatase plays a major role in the calcium-dependent regulation of flagellar motility.