Antimicrobial and Antibiofilm Activities of Helical Antimicrobial Peptide Sequences Incorporating Metal-Binding Motifs
Antimicrobial and Antibiofilm Activities of Helical Antimicrobial Peptide Sequences Incorporating Metal-Binding Motifs
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DOI：
10.1021/acs.biochem.9b00440
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发表时间：
2019-09-10
期刊：
影响因子：
2.9
通讯作者：
Franco, Octavio L.
中科院分区：
文献类型：
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作者：
Agbale, Caleb M.;Sarfo, Justice K.;Franco, Octavio L.
抗菌肽（AMPs）代表了对抗抗生素耐药性这一全球健康问题的替代战略。然而，天然存在的抗菌肽通常不足以作为抗生素使用。需要结合额外设计原则的优化合成版本。本研究将能增强生物功能的氨基末端Cu（II）和Ni(II) （ATCUN）结合基序设计到CM15和citropin1.1这两个AMPs的天然序列中。金属结合基元的结合调节了合成肽对碳青霉烯耐药肠球菌（CRE）细菌的抗菌活性，包括碳青霉烯耐药肺炎克雷伯菌（KpC+）和大肠杆菌（KpC+）。活性调制取决于所使用的ATCUN变体的类型。膜渗透性实验显示，硅选择的铅模板CM15及其ATCUN类似物增加了细菌细胞死亡。质谱分析、圆二色性和分子动力学模拟表明，ATCUN衍生物与Cu（II）离子配位并没有增加AMPS的螺旋倾向。当CM15 ATCUN变体与美罗培南、链霉素或氯霉素联合使用时，显示出对大肠杆菌（KpC+ 1812446）生物膜的协同作用。Motif的添加也降低了野生型AMP的溶血活性，提高了小鼠在全身感染模型中的存活率。这些生物活性依赖于ATCUN基序的特定氨基酸，强调了可能使用大小，电荷和疏水性来微调AMP的生物学功能。我们的数据表明，将金属结合基序结合到肽序列中会导致具有修改生物学特性的合成变体。这些原理可以应用于增强其他肽序列的活性。
Antimicrobial peptides (AMPs) rep resent alternative strategies to combat the global health problem of antibiotic resistance. However, naturally occurring AMPs are generally not sufficiently active for use as antibiotics. Optimized synthetic versions incorporating additional design principles are needed. Here, we engineered amino-terminal Cu(II) and Ni(II) (ATCUN) binding motifs, which can enhance biological function, into the native sequence of two AMPs, CM15 and citropin1.1. The incorporation of metalbinding motifs modulated the antimicrobial activity of synthetic peptides against a panel of carbapenem-resistant enterococci (CRE) bacteria, including carbapenem-resistant Klebsiella pneumoniae (KpC+) and Escherichia coli (KpC+). Activity modulation depended on the type of ATCUN variant utilized. Membrane permeability assays revealed that the in silico selected lead template, CM15, and its ATCUN analogs increased bacterial cell death. Mass spectrometry, circular dichroism, and molecular dynamics simulations indicated that coordinating ATCUN derivatives with Cu(II) ions did not increase the helical tendencies of the AMPS. CM15 ATCUN variants, when combined with Meropenem, streptomycin, or chloramphenicol, showed synergistic effects against E. coli (KpC+ 1812446) biofilms. Motif addition also reduced the hemolytic activity of the wild-type AMP and improved the survival rate of mice in a systemic infection model. The dependence of these bioactivities on the particular amino acids of the ATCUN motif highlights the possible use of size, charge, and hydrophobicity to fine-tune AMP biological function. Our data indicate that incorporating metal-binding motifs into peptide sequences leads to synthetic variants with modified biological properties. These principles may be applied to augment the activities of other peptide sequences.