Antimicrobial activity, bactericidal mechanism and LPS-neutralizing activity of the cell-penetrating peptide pVEC and its analogs
Antimicrobial activity, bactericidal mechanism and LPS-neutralizing activity of the cell-penetrating peptide pVEC and its analogs
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DOI：
10.1002/psc.1408
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发表时间：
2011-12-01
影响因子：
2.1
通讯作者：
Shin, Song Yub
中科院分区：
文献类型：
--
作者：
Nan, Yong Hai;Park, Il-Seon;Shin, Song Yub
pVEC是来源于鼠血管内皮钙粘蛋白的细胞穿透肽。为了评价pVEC作为抗菌肽（AMP）的潜力，我们合成了pVEC及其Trp和Arg/Lys取代的类似物，并研究了它们的抗菌活性和脂多糖（LPS）中和活性。pVEC及其类似物显示出对革兰氏阳性和革兰氏阴性细菌的有效的抗微生物活性（最小抑制浓度：4-16 μ M），但即使在200 μ M的浓度下也没有溶血活性或具有较小的溶血活性（小于10%溶血）。这些肽在4 μ M时诱导对金黄色葡萄球菌的接近完全的膜去极化（超过80%），并且在低至1 μ M的浓度下从细菌膜模拟脂质体中显著的染料渗漏（35-70%）。pVEC及其类似物在脂质体染料渗漏和膜去极化中的荧光分布与蛙源AMP，爪蟾抗菌肽2的荧光分布相似。这些结果表明pVEC及其类似物通过在细胞质膜中形成孔或离子通道来杀死细菌。pVEC及其类似物在10至50 μ M时显著抑制LPS刺激的小鼠巨噬细胞RAW 264.7细胞中一氧化氮的产生或肿瘤坏死因子-α的释放，其中RAW 264.7未被损伤。综上所述，我们的研究结果表明，pVEC及其类似物具有有效的抗菌和LPS中和活性，可以作为抗菌肽用于治疗微生物感染和脓毒症。版权所有（C）2011欧洲肽协会和约翰威利父子有限公司。
pVEC is a cell-penetrating peptide derived from the murine vascular endothelial-cadherin protein. To evaluate the potential of pVEC as antimicrobial peptide (AMP), we synthesized pVEC and its analogs with Trp and Arg/Lys substitution, and their antimicrobial and lipopolysaccharide (LPS)-neutralizing activities were investigated. pVEC and its analogs displayed a potent antimicrobial activity (minimal inhibitory concentration: 4-16 mu M) against Gram-positive and Gram-negative bacteria but no or less hemolytic activity (less than 10% hemolysis) even at a concentration of 200 mu M. These peptides induced a near-complete membrane depolarization (more than 80%) at 4 mu M against Staphylococcus aureus and a significant dye leakage (35-70%) from bacterial membrane-mimicking liposome at a concentration as low as 1 mu M. The fluorescence profiles of pVEC and its analogs in dye leakage from liposome and membrane depolarization were similar to those of a frog-derived AMP, magainin 2. These results suggest that pVEC and its analogs kill bacteria by forming a pore or ion channel in the cytoplasmic membrane. pVEC and its analogs significantly inhibited nitric oxide production or tumor necrosis factor-alpha release in LPS-stimulated mouse macrophage RAW264.7 cells at 10 to 50 mu M, in which RAW264.7 were not damaged. Taken together, our results suggest that pVEC and its analogs with potent antimicrobial and LPS-neutralizing activities can serve as AMPs for the treatment of microbial infection and sepsis. Copyright (C) 2011 European Peptide Society and John Wiley & Sons, Ltd.