DNA Binding and Molecular Dynamic Studies of Polycyclic Tetramate Macrolactams (PTM) with Potential Anticancer Activity Isolated from a Sponge-Associated Streptomyces zhaozhouensis subsp. mycale subsp. nov.
DNA Binding and Molecular Dynamic Studies of Polycyclic Tetramate Macrolactams (PTM) with Potential Anticancer Activity Isolated from a Sponge-Associated Streptomyces zhaozhouensis subsp. mycale subsp. nov.
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DOI：
10.1007/s10126-018-9866-9
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发表时间：
2019-02-01
影响因子：
3
通讯作者：
Sajeevan, T. P.
中科院分区：
文献类型：
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作者：
Dhaneesha, M.;Hasin, O.;Sajeevan, T. P.
从海洋海绵Mycale sp.中分离到一株海绵伴生放线菌(菌株MCCB267)。在印度东南海岸外的印度洋采集。用16S rRNA基因测序对该菌株进行了系统发育研究，结果表明该菌株与肇州链霉菌具有较高的序列相似性。而菌株MCCB267表现出明显的生理生化特征，被命名为肇州链霉菌亚种。麦卡儿。亚种。11月对菌株MCCB267的乙酸乙酯粗提物进行细胞毒性引导分级，得到4个属于多环四酸大内酰胺(PTM)天然产物家族的纯化合物：伊卡鲁霉素(IK)(1)、克利福那胺A(CF)(2)、30-氧代-28-N-甲基力克(OI)(3)和28-N-甲基力克(MI)(4)。这四个化合物在体外对NCI-H460肺癌细胞表现出良好的细胞毒活性，通过诱导细胞凋亡来诱导细胞死亡。流式细胞仪分析显示，1、3和4诱导NCI-H460细胞周期停滞于G1期，而2诱导细胞周期停滞于S期。在用化合物1-4处理癌细胞系时，也检测到处于亚G1期的细胞的浓度依赖积累。体外细胞毒性研究通过分子对接和分子动力学模拟分析得到支持。一项电子计算机研究表明，PTMS可以结合到DNA的小沟上，随后诱导细胞凋亡刺激，导致细胞死亡。本文描述了分离的放线菌的特征，四种PTM，1-4的作用方式的研究，以及分子对接和分子动力学模拟分析。
A sponge-associated actinomycete (strain MCCB267) was isolated from a marine sponge Mycale sp. collected in the Indian Ocean off the Southeast coast of India. Phylogenetic studies of this strain using 16S rRNA gene sequencing showed high sequence similarity to Streptomyces zhaozhouensis. However, strain MCCB267 showed distinct physiological and biochemical characteristic features and was thus designated as S. zhaozhouensis subsp. mycale. subsp. nov. A cytotoxicity-guided fractionation of the crude ethyl acetate extract of strain MCCB267 culture medium yielded four pure compounds belonging to the polycyclic tetramate macrolactam (PTM) family of natural products: ikarugamycin (IK) (1), clifednamide A (CF) (2), 30-oxo-28-N-methylikarugamycin (OI) (3), and 28-N-methylikarugamycin (MI) (4). The four compounds exhibited promising cytotoxic activity against NCI-H460 lung carcinoma cells in vitro, by inducing cell death via apoptosis. Flow cytometric analysis revealed that 1, 3, and 4 induced cell cycle arrest during G1 phase in the NCI-H460 cell line, whereas 2 induced cell arrest in the S phase. A concentration-dependent accumulation of cells in the sub-G1 phase was also detected upon treatment of the cancer cell line with compounds 1-4. The in vitro cytotoxicity studies were supported by molecular docking and molecular dynamic simulation analyses. An in silico study revealed that the PTMs can bind to the minor groove of DNA and subsequently induce the apoptotic stimuli leading to cell death. The characterization of the isolated actinomycete, the study of the mode of action of the four PTMs, 1-4, and the molecular docking and molecular dynamic simulations analyses are herein described.