Mechanistic Study of Membrane Disruption by Antimicrobial Methacrylate Random Copolymers by the Single Giant Vesicle Method
Mechanistic Study of Membrane Disruption by Antimicrobial Methacrylate Random Copolymers by the Single Giant Vesicle Method
复制标题
单巨囊泡法研究抗菌甲基丙烯酸酯无规共聚物膜破坏机理
DOI：
10.1021/acs.langmuir.1c01047
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发表时间：
2021
期刊：
影响因子：
3.9
通讯作者：
Yasuhara, Kazuma
中科院分区：
文献类型：
--
作者：
Tsukamoto, Manami;Zappala, Emanuele;Caputo, Gregory A.;Kikuchi, Jun-ichi;Najarian, Kayvan;Kuroda, Kenichi;Yasuhara, Kazuma
阳离子两亲聚合物已经成为创造通过破坏细菌细胞膜起作用的新的抗微生物材料的平台。虽然活性表征和化学优化已经在许多研究中完成，但我们对聚合物的抗菌机制的了解仍然存在差距，这是连接其化学结构和生物活性所必需的。为此，我们使用了一个单一的巨大的单层囊泡（GUV）的方法来确定甲基丙烯酸酯无规共聚物的膜破坏机制。共聚物由甲基丙烯酸氨乙酯和甲基丙烯酸甲酯（MMA）或甲基丙烯酸丁酯（BMA）的无规序列组成，分子量为1600-2100 g·mol-1。制备由1-棕榈酰基-2-油酰基-sn-甘油-3-磷酸乙醇胺（POPE）和1-棕榈酰基-2-油酰基-sn-甘油-3-磷酸-（1′-rac-甘油）钠盐（POPG）的8：2混合物组成的GUV和仅含有1-棕榈酰基-2-油酰基-sn-甘油-3-磷酸胆碱（POPC）的GUV，以分别模拟细菌（大肠杆菌）或哺乳动物膜。GUV中细菌和哺乳动物细胞膜模拟脂质双层的破坏反映了共聚物的抗微生物和溶血活性，表明共聚物通过破坏细胞膜起作用。与BMA的共聚物在脂质双层中形成孔，而与MMA的共聚物导致GUV破裂。因此，我们提出，该机制是固有的疏水基团的化学身份或属性。与MMA的共聚物表现出特征的S形曲线的GUV突发的时间过程。我们提出了一个新的动力学模型与正反馈回路的聚合物链的脂质双层插入。烷基依赖性膜破坏机制的新发现将为疏水基团在抗菌活性和选择性的优化策略中的作用提供新的见解。
Cationic amphiphilic polymers have been a platform to create new antimicrobial materials that act by disrupting bacterial cell membranes. While activity characterization and chemical optimization have been done in numerous studies, there remains a gap in our knowledge on the antimicrobial mechanisms of the polymers, which is needed to connect their chemical structures and biological activities. To that end, we used a single giant unilamellar vesicle (GUV) method to identify the membrane-disrupting mechanism of methacrylate random copolymers. The copolymers consist of random sequences of aminoethyl methacrylate and methyl (MMA) or butyl (BMA) methacrylate, with low molecular weights of 1600–2100 g·mol–1. GUVs consisting of an 8:2 mixture of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine (POPE) and 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-(1′-rac-glycerol), sodium salt (POPG) and those with only 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) were prepared to mimic the bacterial (Escherichia coli) or mammalian membranes, respectively. The disruption of bacteria and mammalian cell membrane-mimetic lipid bilayers in GUVs reflected the antimicrobial and hemolytic activities of the copolymers, suggesting that the copolymers act by disrupting cell membranes. The copolymer with BMA formed pores in the lipid bilayer, while that with MMA caused GUVs to burst. Therefore, we propose that the mechanism is inherent to the chemical identity or properties of hydrophobic groups. The copolymer with MMA showed characteristic sigmoid curves of the time course of GUV burst. We propose a new kinetic model with a positive feedback loop in the insertion of the polymer chains in the lipid bilayer. The novel finding of alkyl-dependent membrane-disrupting mechanisms will provide a new insight into the role of hydrophobic groups in the optimization strategy for antimicrobial activity and selectivity.