Identification of human sperm proteins that interact with human zona pellucida3 (ZP3) using yeast two-hybrid system.
Identification of human sperm proteins that interact with human zona pellucida3 (ZP3) using yeast two-hybrid system.
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DOI：
10.1016/j.jri.2009.10.006
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发表时间：
2010-01
影响因子：
3.4
通讯作者：
Dhandapani, Latha
中科院分区：
文献类型：
--
作者：
Naz, Rajesh K.;Dhandapani, Latha
与透明带3 （ZP3）相互作用的精子蛋白在人类中尚未被明确鉴定。在本研究中，酵母双杂交（Y2H）系统被用于鉴定与人类ZP3相互作用的人类精子蛋白。将人ZP3 cDNA克隆到pAS2-1酵母载体中，作为诱饵在人睾丸cDNA文库中寻找反应蛋白。获得了6个特异性克隆，进一步证实了它们在哺乳动物双杂交系统中的相互作用。这6个克隆与GenBank数据库中的几个蛋白具有同源性。其中，最强的ZP3相互作用蛋白，与泛素相关蛋白-2样（UBAP2L）具有97%的同源性，在半区试验中被测试。UBAP2L抗体显著（p < 0.001）抑制人精子带结合。我们得出结论，Y2H系统是鉴定与ZP蛋白相互作用的新基因编码蛋白的有用策略。据我们所知，这是首次使用Y2H系统鉴定与人类卵母细胞ZP3相互作用的精子蛋白的研究。使用该系统鉴定的新蛋白可能在阐明受精级联，新一代非甾体避孕药的开发以及人类不孕症的特定诊断和治疗方面找到应用。
Sperm proteins that interact with zona pellucida 3 (ZP3) have not been clearly identified in humans. In the present study, the yeast two-hybrid (Y2H) system was used to identify human sperm proteins that interact with human ZP3. Human ZP3 cDNA was cloned into pAS2-1 yeast vector and used as bait to find reactive proteins in the human testis cDNA library. Six specific clones were obtained that were further confirmed for interaction using the mammalian two-hybrid system. These six clones showed homologies with several proteins in the GenBank database. Of these, the strongest ZP3 interacting protein, that shows 97% homology with ubiquitin associated protein-2 like (UBAP2L), was tested in the hemizona assay. UBAP2L antibodies significantly (p < 0.001) inhibited human sperm-zona binding in this assay. We conclude that the Y2H system is a useful strategy for identifying novel genes encoding proteins that interact with ZP proteins. To our knowledge, this is the first study using the Y2H system to identify sperm proteins that interact with human oocyte ZP3. Novel proteins identified using this system may find applications in elucidating the fertilization cascade, development of a new generation of non-steroidal contraceptives, and specific diagnosis and treatment of human infertility.
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DOI：
10.1083/jcb.123.6.1441
发表时间：
1993-12
期刊：
The Journal of cell biology
影响因子：
--
作者：
Vanderhaeghen P;Schurmans S;Vassart G;Parmentier M
通讯作者：
Parmentier M
影响因子：
3.6
作者：
Sutovsky, P;Manandhar, G;Day, BN
通讯作者：
Day, BN
影响因子：
--
作者：
Spehr, M;Hatt, H
通讯作者：
Hatt, H
影响因子：
1.6
作者：
Harris, JD;Seid, CA;Liu, HF
通讯作者：
Liu, HF
影响因子：
6.1
作者：
Samuel, A. S.;Naz, R. K.
通讯作者：
Naz, R. K.