Synthesis, characterization, antimicrobial activity and LPS-interaction properties of SB041, a novel dendrimeric peptide with antimicrobial properties
Synthesis, characterization, antimicrobial activity and LPS-interaction properties of SB041, a novel dendrimeric peptide with antimicrobial properties
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DOI：
10.1016/j.peptides.2010.04.022
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发表时间：
2010-08-01
期刊：
影响因子：
3
通讯作者：
Rinaldi, Andrea C.
中科院分区：
文献类型：
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作者：
Bruschi, Michela;Pirri, Giovanna;Rinaldi, Andrea C.
与单体肽相比，多聚肽具有多种优势，例如肽酶和蛋白酶的活性更高、稳定性更高。 SB041是一种具有树枝状结构的新型抗菌肽；它是由赖氨酸核心与氨基戊酸链连接的pyrEKKIRVRLSA 四聚体。在这里，我们报告了其合成、NMR 表征、抗菌活性和 LPS 相互作用特性。该肽对革兰氏阴性菌株特别有效，其效力（以摩尔计）与脂肽粘菌素和多粘菌素 B 相当，但它也对选定的革兰氏阳性菌株表现出一定的活性。根据这些迹象，我们研究了 SB041 在体外结合大肠杆菌和铜绿假单胞菌 LPS 的功效，并抵消其在源自 RAW 264.7 巨噬细胞的 RAW-Blue (TM) m 细胞中的生物效应。荧光置换试验证明，SB041 与纯化的 LPS，尤其是大肠杆菌的 LPS 强烈结合，并且很容易渗透到 LPS 单层中。然而，SB041对大肠杆菌的杀灭活性并没有因为培养基中LPS浓度的增加而受到抑制。检查 SB041 对 Raw-Blue 细胞中 LPS 诱导的模式识别受体 (PRR) 激活的影响发现，虽然当 RAW-Blue 细胞受到铜绿假单胞菌 LPS 攻击时，该肽在统计学上显着降低了 PRR 刺激，但当使用大肠杆菌 LPS 激活先天免疫防御样反应时，没有观察到相同情况。因此，正如之前对其他抗菌肽所观察到的，SB041 与 LPS 的结合也不一定转化为 LPS 中和活性，这表明 SB041-LPS 相互作用必定具有复杂的性质。 (C) 2010 Elsevier Inc. 保留所有权利。
Multimeric peptides offer several advantages with respect to their monomeric counterparts, as increased activity and greater stability to peptidases and proteases. SB041 is a novel antimicrobial peptide with dendrimeric structure; it is a tetramer of pyrEKKIRVRLSA linked by a lysine core, with an amino valeric acid chain. Here, we report on its synthesis, NMR characterization, antimicrobial activity, and LPS-interaction properties. The peptide was especially active against Gram-negative strains, with a potency comparable (on molar basis) to that of lipopeptides colistin and polymixin B, but it also displayed some activity against selected Gram-positive strains. Following these indications, we investigated the efficacy of SB041 in binding Escherichia coli and Pseudomonas aeruginosa LPS in vitro and counteracting its biological effects in RAW-Blue (TM) m cells, derived from RAW 264.7 macrophages. SB041 strongly bound purified LPS, especially that of E. coli, as proved by fluorescent displacement assay, and readily penetrated into LPS monolayers. However, the killing activity of SB041 against E. coli was not inhibited by increasing concentrations of LPS added to the medium. Checking the SB041 effect on LPS-induced activation of pattern recognition receptors (PRRs) in Raw-Blue cells revealed that while the peptide gave a statistically significant decrease in PRRs stimulation when RAW-Blue cells were challenged with P. aeruginosa LPS, the same was not seen when E. coli LPS was used to activate innate immune defense-like responses. Thus, as previously seen for other antimicrobial peptides, also for SB041 binding to LPS did not translate necessarily into LPS-neutralizing activity, suggesting that SB041-LPS interactions must be of complex nature. (C) 2010 Elsevier Inc. All rights reserved.