Actiflagelin, a new sperm activator isolated from Walterinnesia aegyptia venom using phenotypic screening.
Actiflagelin, a new sperm activator isolated from Walterinnesia aegyptia venom using phenotypic screening.
复制标题
Actiflagelin是一种使用表型筛选从Walterinnesia Aegyptia venom分离出的新精子激活剂。
DOI：
10.1186/s40409-018-0140-4
复制
发表时间：
2018
期刊：
影响因子：
--
通讯作者：
De Waard M
中科院分区：
文献类型：
--
作者：
Abd El-Aziz TM;Al Khoury S;Jaquillard L;Triquigneaux M;Martinez G;Bourgoin-Voillard S;Sève M;Arnoult C;Beroud R;De Waard M
精子含有丰富的细胞表面受体和离子通道，这是其大多数基本功能（如运动和顶体反应）所必需的。相反，动物毒液富含生物活性化合物，主要针对这些离子通道和细胞表面受体。因此，我们假设，动物毒液中应该含有足够丰富的精子调节化合物，可以用于药物发现项目。我们的目的是通过使用基于精子的表型筛选来鉴定埃及Walterinnesia aegyptia毒液的阳性调节剂来证明这一事实。在此，为了证明毒液中含有对精子生理有益的化合物这一概念，我们用反相高效液相色谱法（RP-HPLC）分离了埃及Walterinnesia aegyptia蛇毒，并筛选了能够加速小鼠精子运动的生物活性组分（初步筛选）。接下来，我们通过阳离子交换从阳性部分中纯化每个化合物，并通过二次筛选鉴定生物活性肽。通过对还原/烷基化化合物进行Edman测序，并结合胰蛋白酶或V8蛋白酶酶切后还原/烷基化片段肽的LC-ESI-QTOF MS/MS分析，建立肽序列。利用这种两步纯化方案结合细胞表型筛选，我们从OF1雄性小鼠中鉴定出了一种新的毒素7329.38 Da（激活鞭毛素），可以激活体外精子活力。Actiflagelin的长度为63个氨基酸，包含五个二硫桥，沿二硫连接模式C1-C5， C2-C3, C4-C6， C7-C8和C9-C10。对其结构的建模表明，它属于三指毒素家族，与布旦丁有明显的同源性，布旦丁是一种来自白蜡毒的肽。本报告证明了鉴定生育化合物的可行性，这可能是治疗不孕不育的情况下，运动是一个问题的潜力。
Sperm contains a wealth of cell surface receptors and ion channels that are required for most of its basic functions such as motility and acrosome reaction. Conversely, animal venoms are enriched in bioactive compounds that primarily target those ion channels and cell surface receptors. We hypothesized, therefore, that animal venoms should be rich enough in sperm-modulating compounds for a drug discovery program. Our objective was to demonstrate this fact by using a sperm-based phenotypic screening to identify positive modulators from the venom of Walterinnesia aegyptia. Herein, as proof of concept that venoms contain interesting compounds for sperm physiology, we fractionated Walterinnesia aegyptia snake venom by RP-HPLC and screened for bioactive fractions capable of accelerating mouse sperm motility (primary screening). Next, we purified each compound from the positive fraction by cation exchange and identified the bioactive peptide by secondary screening. The peptide sequence was established by Edman sequencing of the reduced/alkylated compound combined to LC-ESI-QTOF MS/MS analyses of reduced/alkylated fragment peptides following trypsin or V8 protease digestion. Using this two-step purification protocol combined to cell phenotypic screening, we identified a new toxin of 7329.38 Da (actiflagelin) that activates sperm motility in vitro from OF1 male mice. Actiflagelin is 63 amino acids in length and contains five disulfide bridges along the proposed pattern of disulfide connectivity C1-C5, C2-C3, C4-C6, C7-C8 and C9-C10. Modeling of its structure suggests that it belongs to the family of three finger toxins with a noticeable homology with bucandin, a peptide from Bungarus candidus venom. This report demonstrates the feasibility of identifying profertility compounds that may be of therapeutic potential for infertility cases where motility is an issue.
登录
查看更多内容
影响因子：
2.9
作者：
El Aziz, Tarek Mohamed Abd;Bourgoin-Voillard, Sandrine;De Waard, Michel
通讯作者：
De Waard, Michel
影响因子：
4.5
作者：
Badr G;Al-Sadoon MK;El-Toni AM;Daghestani M
通讯作者：
Daghestani M
影响因子：
2.8
作者：
Carbajal-Saucedo, Alejandro;Lopez-Vera, Estuardo;Olvera-Rodriguez, Alejandro
通讯作者：
Olvera-Rodriguez, Alejandro
影响因子：
4
作者：
Martinez, Guillaume;Hograindleur, Jean-Pascal;Arnoult, Christophe
通讯作者：
Arnoult, Christophe
影响因子：
2.8
作者：
LEE, CY;CHEN, YM;MEBS, D
通讯作者：
MEBS, D