Macromolecular interactions of triterpenoids and targeted toxins: role of saponins charge.
Macromolecular interactions of triterpenoids and targeted toxins: role of saponins charge.
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DOI：
10.1016/j.ijbiomac.2013.07.008
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发表时间：
2013-10
影响因子：
8.2
通讯作者：
M. Thakur;A. Weng;Alexandra Pieper;K. Mergel;Benedicta von Mallinckrodt;R. Gilabert-Oriol;C. Görick;B. Wiesner;Jenny Eichhorst;M. Melzig;H. Fuchs
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文献类型：
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作者：
M. Thakur;A. Weng;Alexandra Pieper;K. Mergel;Benedicta von Mallinckrodt;R. Gilabert-Oriol;C. Görick;B. Wiesner;Jenny Eichhorst;M. Melzig;H. Fuchs
蛋白毒素与某些植物三萜类化合物的大分子相互作用在肿瘤治疗中具有潜在的应用价值。评价了仅某些皂苷增强肿瘤细胞中特异性毒素的内体逃逸的能力，并将其与电泳迁移率相关联。本文以Saponaria officinalisLinn的皂苷为先导，详细研究了这一进化保守的原理。利用琼脂糖凝胶电泳获得具有不同电泳迁移率的纯皂苷级分，通过活细胞监测测试其增强毒性的能力。分离出相对电泳迁移率为（-0.05，0.41，0.59，0.75和1.00）的五个级分（SOG 1-SOG 5），并使用薄层色谱法、HPLC和质谱分析进行评价。细胞毒性实验显示使用SOG 3的最高有效性。用SOG 3进行的活细胞成像实验显示，这种具有0.59的特定REM的皂苷可以辅助毒性有效负载的溶酶体/内体释放，而不影响质膜的完整性，并且可以导致诱导细胞凋亡。与细菌毒素相比，这种电荷依赖性增强也被发现对I型核糖体失活蛋白具有高度特异性。植物毒素和皂苷与肿瘤细胞的电荷相互作用在毒素特异性调节反应中起主要作用。这一发现开辟了新的方法，发现蛋白质皂苷相互作用的进化保守，并测试他们的作用，治疗分子的内体逃逸。
Macromolecular interaction of protein toxins with certain plant triterpenoids holds potential for application in tumor therapy. The ability of only certain saponins to enhance the endosomal escape of toxins specifically in tumor cells was evaluated and set into correlation with the electrophoretic mobility. Saponins fromSaponaria officinalisLinn, were selected as a lead to understand this evolutionarily conserved principle in detail. Agarose gel electrophoresis was utilized to procure pure saponin fractions with different electrophoretic mobility, which were tested for their ability to enhance the toxicity by live cell monitoring. Five fractions (SOG1–SOG5) were isolated with a relative electrophoretic mobility of (−0.05, 0.41, 0.59, 0.75 and 1.00) and evaluated using thin layer chromatography, HPLC, and mass spectroscopic analysis. Cytotoxicity experiments revealed highest effectiveness with SOG3. Live cell imaging experiments with SOG3 revealed that this saponin with a specific REM of 0.59 could assist in the lyso/endosomal release of the toxic payload without affecting the integrity of plasma membrane and could lead to the induction of apoptosis. This charge dependent enhancement was also found to be highly specific to type I ribosome inactivating proteins compared to bacterial toxins.Charge interaction of plant toxins and saponins with tumor cells, plays a major role in toxin specific modulation of response. The finding opens up newer ways of finding protein saponin interaction conserved evolutionarily and to test their role in endosomal escape of therapeutic molecules.