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首页> 外文期刊>Developmental and Comparative Immunology: Ontogeny, Phylogeny, Aging: The Official Journal of the International Society of Developmental and Comparative Immunology >Identification and functional characterization of an uncharacterized antimicrobial peptide from a ciliate Paramecium caudatum
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Identification and functional characterization of an uncharacterized antimicrobial peptide from a ciliate Paramecium caudatum

机译：纤毛虫草履虫未鉴定的抗菌肽的鉴定和功能表征

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摘要

The global ever-growing concerns about multi-drug resistant (MDR) microbes leads to urgent demands for exploration of new antibiotics including antimicrobial peptides (AMPs). Here we demonstrated that a cDNA from Ciliata Paramecium caudatum, designated Pcamp1, coded for a protein with features characteristic of AMPs, which is not homologous to any AMPs currently known. Both the C-terminal 91 amino acid residues of PcAMP1, cPcAMP1, expressed in Escherichia coli and the C-terminal 26 amino acid residues (predicted mature AMP), cPcAMP1/26, synthesized, underwent a coil-to-helix transition in the presence of TFE, SDS or DPC. Functional assays revealed that cPcAMP1 and cPcAMP1/26 were both able to kill Aeromonas hydrophila and Staphylococcus aureus. ELISA showed that cPcAMP1 and cPcAMP1/26 were able to bind to microbe-associated molecular pattern molecules LPS and LTA, which was further corroborated by the observations that cPcAMP1 could deposit onto the bacterial membranes. Importantly, both cPcAMP1 and cPcAMP1/26 were able to induce bacterial membrane permeabilization and depolarization, and to increase intracellular ROS levels. Additionally, cPcAMP1 and cPcAMP1/26 were not cytotoxic to mammalian cells. Taken together, our results show that PcAMP1 is a potential AMP with a membrane selectivity towards bacterial cells, which renders it a promising template for the design of novel peptide antibiotics against MDR microbes. It also shows that use of signal conserved sequence of AMPs can be an effective tool to identify potential AMPs across different animal classes. (C) 2016 Elsevier Ltd. All rights reserved.
机译：全球对耐多药（MDR）微生物的关注日益增加，因此迫切需要探索包括抗菌肽（AMPs）在内的新抗生素。在这里，我们证明了来自尾纤毛虫的CcDNA，称为Pcamp1，编码具有AMPs特征的蛋白，该蛋白与目前已知的AMPs不同源。在大肠杆菌中表达的PcAMP1，cPcAMP1的C末端91个氨基酸残基和合成的c末端26个氨基酸残基（预测的成熟AMP）cPcAMP1 / 26均在存在下从线圈向螺旋过渡TFE，SDS或DPC。功能测定表明，cPcAMP1和cPcAMP1 / 26均能够杀死嗜水气单胞菌和金黄色葡萄球菌。酶联免疫吸附测定表明cPcAMP1和cPcAMP1 / 26能够与微生物相关的分子模式分子LPS和LTA结合，这被cPcAMP1可以沉积到细菌膜上的观察进一步证实。重要的是，cPcAMP1和cPcAMP1 / 26均能够诱导细菌膜通透和去极化，并能增加细胞内ROS水平。此外，cPcAMP1和cPcAMP1 / 26对哺乳动物细胞没有细胞毒性。两者合计，我们的结果表明PcAMP1是潜在的AMP，对细菌细胞具有膜选择性，这使其成为设计针对MDR微生物的新型肽抗生素的有希望的模板。它还表明，使用信号保守的AMP序列可以是一种有效的工具，可以识别不同动物类别中潜在的AMP。 （C）2016 Elsevier Ltd.保留所有权利。

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