Identification and functional characterization of an uncharacterized antimicrobial peptide from a ciliate Paramecium caudatum
Identification and functional characterization of an uncharacterized antimicrobial peptide from a ciliate Paramecium caudatum
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DOI：
10.1016/j.dci.2016.02.016
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发表时间：
2016-07-01
影响因子：
2.9
通讯作者：
Zhang, Shicui
中科院分区：
文献类型：
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作者：
Cui, Pengfei;Dong, Yuan;Zhang, Shicui
全球对多重耐药（MDR）微生物的日益关注导致迫切需要开发包括抗菌肽（AMP）在内的新抗生素。在这里，我们表明，从纤毛虫草履虫尾，命名为Pcamp 1，编码的蛋白质与AMP的特征，这是不同源的任何AMP目前已知的。在TFE、SDS或DPC的存在下，在大肠杆菌中表达的PcAMP 1的C-末端91个氨基酸残基（cPcAMP 1）和合成的C-末端26个氨基酸残基（预测成熟AMP）（cPcAMP 1/26）都经历了螺旋到螺旋的转变。功能试验表明，cPcAMP 1和cPcAMP 1/26均能杀灭嗜水气单胞菌和金黄色葡萄球菌。酶联免疫吸附试验（ELISA）结果表明，cPcAMP 1和cPcAMP 1/26能与细菌相关分子模式分子LPS和LTA结合，并能在细菌细胞膜上存款。重要的是，cPcAMP 1和cPcAMP 1/26都能够诱导细菌膜透化和去极化，并增加细胞内ROS水平。此外，cPcAMP 1和cPcAMP 1/26对哺乳动物细胞无细胞毒性。总之，我们的研究结果表明，PcAMP 1是一个潜在的AMP与膜选择性对细菌细胞，这使得它成为一个有前途的模板设计的新的肽抗生素对MDR微生物。这也表明，使用AMP的信号保守序列可以是一种有效的工具，以确定潜在的AMP在不同的动物类别。(C)2016爱思唯尔有限公司版权所有
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.