de Souza K R, Nunes L O, Salnikov E S, Mundim H M, Munhoz V H O, Lião L M, Aisenbrey C, Resende J M, Bechinger B, Verly R M
Departamento de Química, Universidade Federal dos Vales do Jequitinhonha e Mucuri, 39100-000 Diamantina, MG, Brazil; Université de Strasbourg/CNRS, UMR7177, Institut de Chimie, 67000 Strasbourg, France.
Departamento de Química, Universidade Federal dos Vales do Jequitinhonha e Mucuri, 39100-000 Diamantina, MG, Brazil.
Biophys Chem. 2025 Feb;317:107353. doi: 10.1016/j.bpc.2024.107353. Epub 2024 Nov 14.
Here we present studies of the structure and membrane interactions of ecPis-4 s, a new antimicrobial peptide from the piscidin family, which shows a wide-range of potential biotechnological applications. In order to understand the mode of action ecPis-4 s, the peptide was chemically synthesized and structural investigations in the presence of anionic POPC:POPG (3:1, mol:mol) membrane and SDS micelles were performed. CD spectroscopy demonstrated that ecPis-4 s has a high content of helical structure in both membrane mimetic media, which is in line with solution NMR spectroscopy that revealed an amphipathic helical conformation throughout the entire peptide chain. Solid-state NMR experiments of ecPis-4 s selectively labeled with N/H and reconstituted into uniaxially oriented POPC:POPG membranes revealed an ideal partition of hydrophilic and hydrophobic residues within the bilayer interface. The peptide aligns in parallel to the membrane surface, a topology stabilized by aromatic side-chain interactions of the Phe-1, Phe-2 and Trp-9 with the phospholipids. H NMR experiments using deuterated lipids revealed that anionic lipid accumulates in the vicinity of the cationic peptide upon peptide-membrane binding.
在此，我们展示了对ecPis-4 s的结构及其与膜相互作用的研究，ecPis-4 s是一种来自杀鱼毒素家族的新型抗菌肽，具有广泛的潜在生物技术应用。为了了解ecPis-4 s的作用模式，我们对该肽进行了化学合成，并在阴离子型POPC:POPG（3:1，摩尔比）膜和SDS胶束存在的情况下进行了结构研究。圆二色光谱表明，ecPis-4 s在两种膜模拟介质中都具有高含量的螺旋结构，这与溶液核磁共振光谱结果一致，后者显示整个肽链呈两亲性螺旋构象。用N/H选择性标记并重构到单轴取向的POPC:POPG膜中的ecPis-4 s的固态核磁共振实验表明，亲水和疏水残基在双层界面内实现了理想分布。该肽与膜表面平行排列，这种拓扑结构通过Phe-1、Phe-2和Trp-9的芳香族侧链与磷脂的相互作用得以稳定。使用氘代脂质的氢核磁共振实验表明，肽与膜结合时，阴离子脂质会在阳离子肽附近聚集。
