Alteramide B is a microtubule antagonist of inhibiting Candida albicans.
Alteramide B is a microtubule antagonist of inhibiting Candida albicans.
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DOI：
10.1016/j.bbagen.2016.06.025
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发表时间：
2016-10
期刊：
影响因子：
--
通讯作者：
Du L
中科院分区：
文献类型：
--
作者：
Ding Y;Li Y;Li Z;Zhang J;Lu C;Wang H;Shen Y;Du L
交替酰胺B（AT B）是从产酶溶杆菌C3中分离得到的一种新的多环四酸酯大环内酰胺（PTM）。ATB对几种酵母菌，特别是白色念珠菌SC 5314具有较强的抑制活性，但其抗真菌机制尚不清楚。ATB的结构通过广泛的光谱分析确定，包括高分辨率质谱、1D和2D-NMR以及CD光谱。采用流式细胞术、荧光显微镜、透射电镜、分子模拟、过表达和定点突变等方法研究ATB抗念珠菌的分子机制。ATB诱导C.通过破坏微管诱导活性氧（ROS）的产生来抑制白念珠菌。分子动力学研究揭示了ATB与β-微管蛋白亚基的结合模式。β-tubulin基因野生型和定点突变体（TUBB）的过表达改变了C.白念珠菌与ATB结合，证实ATB与β-微管蛋白结合，并表明结合位点为L215、L217、L273、L274和R282。在体内，ATB显著提高了念珠菌病小鼠的存活率并降低了真菌负荷。ATB诱导C.白念珠菌是通过抑制微管蛋白聚合，导致细胞周期停滞在G2/M期。ATB的鉴定及其活性的研究为PTM对抗人类病原体的作用模式提供了新的机制见解。本研究表明ATB是一种新型的微管抑制剂，是一种很有前途的抗念珠菌先导化合物。这些结果也支持β-微管蛋白作为抗念珠菌药物发现的潜在靶点。ATB是一种新的多环四酸酯大环内酰胺（PTM）化合物，在体外和体内对白色念珠菌SC 5314表现出较强的活性。ATB诱导C.通过破坏微管诱导活性氧（ROS）的产生来抑制白念珠菌。ATB与β-微管蛋白的结合模型由Amber 12模拟并由PyMoL显示。
Alteramide B (ATB), isolated from Lysobacter enzymogenes C3, was a new polycyclic tetramate macrolactam (PTM). ATB exhibited potent inhibitory activity against several yeasts, particularly Candida albicans SC5314, but its antifungal mechanism is unknown. The structure of ATB was established by extensive spectroscopic analyses, including high-resolution mass spectrometry, 1D- and 2D-NMR, and CD spectra. Flow cytometry, fluorescence microscope, transmission electron microscope, molecular modeling, overexpression and site-directed mutation studies were employed to delineate the anti-candida molecular mechanism of ATB. ATB induced apoptosis in C. albicans through inducing reactive oxygen species (ROS) production by disrupting microtubules. Molecular dynamics studies revealed the binding patterns of ATB to the β-tubulin subunit. Overexpression of the wild type and site-directed mutants of the β-tubulin gene (TUBB) changed the sensitivity of C. albicans to ATB, confirming the binding of ATB to β-tubulin, and indicating that the binding sites are L215, L217, L273, L274 and R282. In vivo, ATB significantly improved the survival of the candidiasis mice and reduced fungal burden. The molecular mechanism underlying the ATB-induced apoptosis in C. albicans is through inhibiting tubulin polymerization that leads to cell cycle arrest at the G2/M phase. The identification of ATB and the study of its activity provide novel mechanistic insights into the mode of action of PTMs against the human pathogen. This study shows that ATB is a new microtubule inhibitor and a promising anti-Candida lead compound. The results also support β-tubulin as a potential target for anti-Candida drug discovery. ATB, a new polycyclic tetramate macrolactam (PTM) compound, exhibited potent activity against Candida albicans SC5314 in vitro and in vivo. ATB induced apoptosis of C. albicans through inducing the production of reactive oxygen species (ROS) by disrupting microtubules. The binding model of ATB to β-tubulin was simulated by Amber12 and shown by PyMoL.