Protein Kinase A Activity And Protein Phosphorylation During The Mouse Sperm Acrosomal Reaction
Protein Kinase A Activity And Protein Phosphorylation During The Mouse Sperm Acrosomal Reaction
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DOI：
10.1080/014850190512743
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发表时间：
2005-01
期刊：
影响因子：
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通讯作者：
N. Kuji;Y. Tanaka;S. Komatsu;Y. Yoshimura
中科院分区：
文献类型：
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作者：
N. Kuji;Y. Tanaka;S. Komatsu;Y. Yoshimura
使用二维凝胶电泳，蛋白磷酸化引起的环核苷酸依赖性蛋白激酶的变化进行了分析，或没有暴露于蛋白激酶抑制剂，H-8，在小鼠精子顶体反应。用二丁酰环磷酸腺苷或二丁酰环磷酸鸟苷处理精子引起的顶体反应可被H-8抑制。精子提取物诱导的环腺苷酸依赖性蛋白激酶（PKA）和环GMP依赖性蛋白激酶的活性也被H-8抑制。当精子中的内源性PKA被激活，通过添加环AMP，45-kDa的蛋白质点，电泳鉴定表明在体内磷酸化的发生。此外，增强的磷酸化的45-kDa的蛋白点被H-8抑制。这些结果表明，PKA催化的45-kDa蛋白的磷酸化可能参与调节小鼠精子顶体反应。
Using two-dimensional gel electrophoresis, changes in protein phosphorylation caused by cyclic nucleotide-dependent protein kinases were analyzed with or without exposure to a protein kinase inhibitor, H-8, during the mouse sperm acrosomal reaction. The acrosomal reaction, induced by the treatment of sperm with dibutyryl cyclic AMP or dibutyryl cyclic GMP, was inhibited by H-8. The activities of cyclic AMP-dependent protein kinase (PKA) and cyclic GMP-dependent protein kinase induced by the sperm extract were also inhibited by H-8. When endogenous PKA in sperm was activated by the addition of cyclic AMP, a 45-kDa protein spot identified by electrophoresis indicated the occurrence of phosphorylation in vivo. Furthermore, the enhanced phosphorylation of the 45-kDa protein spot was inhibited by H-8. These results suggest that the PKA-catalyzed phosphorylation of the 45-kDa protein may be involved in the regulation of the mouse sperm acrosomal reaction.