Park Seong-Cheol, Kim Heabin, Kim Jin-Young, Kim Hyeonseok, Cheong Gang-Won, Lee Jung Ro, Jang Mi-Kyeong
Department of Polymer Science and Engineering, Sunchon National University, Suncheon 57922, Korea.
Division of Applied Life Sciences and Research Institute of Natural Science, Gyeongsang National University, Jinju 52828, Korea.
Antibiotics (Basel). 2020 Dec 18;9(12):921. doi: 10.3390/antibiotics9120921.
Several antimicrobial peptides (AMPs) have been discovered, developed, and purified from natural sources and peptide engineering; however, the clinical applications of these AMPs are limited owing to their lack of abundance and side effects related to cytotoxicity, immunogenicity, and hemolytic activity. Accordingly, to improve cell selectivity for pseudin-2, an AMP from skin, in mammalian cells and pathogenic fungi, the sequence of pseudin-2 was modified by alanine or lysine at each position of two amino acids within the leucine-zipper motif. Alanine-substituted variants were highly selective toward fungi over HaCaT and erythrocytes and maintained their antifungal activities and mode of action (membranolysis). However, the antifungal activities of lysine-substituted peptides were reduced, and the compound could penetrate into fungal cells, followed by induction of mitochondrial reactive oxygen species and cell death. In vivo antifungal assays of analogous peptide showed excellent antifungal efficiency in a skin infection mouse model. Our results demonstrated the usefulness of selective amino acid substitution in the repeated sequence of the leucine-zipper motif for the design of AMPs with potent antimicrobial activities and low toxicity.
人们已经从天然来源和肽工程中发现、开发并纯化了几种抗菌肽（AMPs）；然而，这些抗菌肽的临床应用受到限制，因为它们的数量不足，且存在与细胞毒性、免疫原性和溶血活性相关的副作用。因此，为了提高来自皮肤的抗菌肽pseudin-2在哺乳动物细胞和致病真菌中的细胞选择性，在亮氨酸拉链基序的两个氨基酸的每个位置用丙氨酸或赖氨酸对pseudin-2的序列进行了修饰。丙氨酸取代的变体对真菌的选择性高于HaCaT细胞和红细胞，并保持了它们的抗真菌活性和作用方式（膜溶解）。然而，赖氨酸取代的肽的抗真菌活性降低，并且该化合物可以渗透到真菌细胞中，随后诱导线粒体活性氧和细胞死亡。类似肽的体内抗真菌试验在皮肤感染小鼠模型中显示出优异的抗真菌效率。我们的结果证明了在亮氨酸拉链基序的重复序列中进行选择性氨基酸取代对于设计具有强大抗菌活性和低毒性的抗菌肽的有用性。
