Dye-release assay for investigation of antimicrobial peptide activity in a competitive lipid environment
Dye-release assay for investigation of antimicrobial peptide activity in a competitive lipid environment
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DOI：
10.1007/s00249-014-0970-0
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发表时间：
2014-09-01
影响因子：
2
通讯作者：
Separovic, Frances
中科院分区：
文献类型：
--
作者：
Sani, Marc-Antoine;Gagne, Eve;Separovic, Frances
提出了一种染料释放法，用于研究竞争性脂环境对两种膜破坏抗菌肽(AMP)活性的影响，这两种抗菌肽分别是斑蛋白1.1和金黄色1.2。结果支持一般结论，即AMP对带负电的膜，例如细菌膜，比在真核细胞中发现的中性膜表面具有更大的亲和力，但仅在竞争性脂环境中。事实上，在单一模型膜环境中，这两种多肽对中性小泡的作用比对带电小泡的作用更强。这种方法也被用来研究在中性脂环境中预先孵育多肽，然后引入带电脂泡的效果。Maculatin被证明从已经形成孔的中性脂双层迁移到带电的膜双层。这一结果也在荷电双分子层中观察到，但有趣的是，中性到中性脂质界面没有观察到这个结果。Arein能够从任一种脂质环境中迁移，这表明与脂膜的结合较弱，并且脂双层的裂解有不同的分子机制。竞争性脂质环境可以用来评估调节膜多肽或蛋白质活性的其他关键条件。
A dye-release method for investigating the effect of a competitive lipid environment on the activity of two membrane-disrupting antimicrobial peptides (AMP), maculatin 1.1 and aurein 1.2, is presented. The results support the general conclusion that AMP have greater affinity for negatively charged membranes, for example bacterial membranes, than for the neutral membrane surface found in eukaryotic cells, but only within a competitive lipid environment. Indeed, in a single-model membrane environment, both peptides were more potent against neutral vesicles than against charged vesicles. The approach was also used to investigate the effect of pre-incubating the peptides in a neutral lipid environment then introducing charged lipid vesicles. Maculatin was shown to migrate from the neutral lipid bilayers, where pores had already formed, to the charged membrane bilayers. This result was also observed for charged-to-charged bilayers but, interestingly, not for neutral-to-neutral lipid interfaces. Aurein was able to migrate from either lipid environment, indicating weaker binding to lipid membranes, and a different molecular mechanism for lysis of lipid bilayers. Competitive lipid environments could be used to assess other critical conditions that modulate the activity of membrane peptides or proteins.