Correlation of epididymal protease inhibitor and fibronectin in human semen.
Correlation of epididymal protease inhibitor and fibronectin in human semen.
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人精液中附睾蛋白酶抑制剂与纤连蛋白的相关性
DOI：
10.1371/journal.pone.0082600
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Wang Z
中科院分区：
文献类型：
--
作者：
Zhang X;Fang J;Xu B;Zhang S;Su S;Song Z;Deng Y;Wang H;Zhao D;Niu X;Wang Z
目的附睾蛋白酶抑制剂（Eppin）位于精子表面，调节人类精液的液化。在这里，我们鉴定了 Eppin 的相关蛋白伴侣，以探索人类精液液化的分子机制。方法(1)采用Western blotting将人精囊蛋白转移至膜上，二维电泳分离，并在重组Epin中孵育。通过质谱 (MS) 鉴定相关蛋白 (2)。 Western blotting检测rEppin与rFibronectin(Fn)的关系； (3)免疫荧光检测精子共定位； (4)通过免疫共沉淀证明了Epin和Fn的相关性。结果 MS 鉴定 Fn 为重组 Eppin 的结合配偶体。制备了 Eppin 重组体并证明 Eppin 片段结合 fn 607-1265 片段。 Eppin-Fn 复合物存在于精子尾部，特别是人类射精精子的中段区域。免疫沉淀表明精子裂解物中的 Eppin 与 Fn 形成复合物。结论 我们的研究表明，Eppin 和 Fn 在人类精液和人类射出的精子中相互结合。 Eppin-Fn 复合物可能参与精液凝固、液化以及精子在女性生殖道中的存活和准备以实现生育能力。
Objective Epididymal protease inhibitor (Eppin) was located on the surface of spermatozoa and modulates the liquefaction of human semen. Here, we identify the correlative protein partner of Eppin to explore the molecular mechanism of liquefaction of human semen. Methods (1) Human seminal vesicle proteins were transferred on the membrane by Western blotting and separated by 2-D electrophoresis and incubated in recombinant Eppin. The correlative protein was identified by Mass Spectrometry (MS) (2). Western blotting was used to determine the relation of rEppin and rFibronectin(Fn); (3) Co-localization in spermatozoa were detected using immunofluorescence; (4) Correalation of Eppin and Fn was proved by co-immunoprecipitation. Results Fn was identified as the binding partner of recombinant Eppin by MS. Recombinant of Eppin was made and demonstrated that the Eppin fragment binds the fn 607-1265 fragment. The Eppin-Fn complex presents on the sperm tail and particularly in the midpiece region of human ejaculated spermatozoa. Immunoprecipitation indicated that Eppin in the spermatozoa lysates was complexed with Fn. Conclusions Our study demonstrates that Eppin and Fn bind to each other in human semen and on human ejaculated spermatozoa. Eppin-Fn complex may involve in semen coagulation, liquefaction and the survival and preparation of spermatozoa for fertility in the female reproductive tract.
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影响因子：
2.8
作者：
Fung, KYC;Glode, LM;Duncan, MW
通讯作者：
Duncan, MW
影响因子：
3.8
作者：
Thys, Mirjan;Nauwynck, Hans;Van Soom, Ann
通讯作者：
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影响因子：
2.5
作者：
FUSI, FM;VIGNALI, M;BRONSON, RA
通讯作者：
BRONSON, RA
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作者：
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通讯作者：
LILJA, H
影响因子：
56.9
作者：
O'Rand, MG;Widgren, EE;Rao, AJ
通讯作者：
Rao, AJ