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Identification of a novel cathelicidin antimicrobial peptide from ducks and determination of its functional activity and antibacterial mechanism

机译:鸭中一种新型cathelicidin抗菌肽的鉴定及其功能活性和抗菌机理的确定

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摘要

The family of antimicrobial peptide, cathelicidins, which plays important roles against infections in animals, has been identified from many species. Here, we identified a novel avian cathelicidin ortholog from ducks and named dCATH. The cDNA sequence of dCATH encodes a predicted 146-amino-acid polypeptide composed of a 17-residue signal peptide, a 109-residue conserved cathelin domain and a 20-residue mature peptide. Phylogenetic analysis demonstrated that dCATH is highly divergent from other avian peptides. The α-helical structure of the peptide exerted strong antimicrobial activity against a broad range of bacteria in vitro, with most minimum inhibitory concentrations in the range of 2 to 4 μM. Moreover, dCATH also showed cytotoxicity, lysing 50% of mammalian erythrocytes in the presence or absence of 10% fetal calf serum at concentrations of 32 μM or 20 μM and killing 50% HaCaT cells at a concentration of 10 μM. The effects on bacterial outer and inner membranes, as examined by scanning electron microscope and transmission electron microscopy, indicate that dCATH kills microbial cells by increasing permeability, causing a loss of membrane integrity.
机译:已从许多物种中鉴定出了抗菌肽Cathelicidins家族,它们在对抗动物感染中起着重要作用。在这里,我们从鸭群中鉴定出一种新颖的鸟类cathelicidin ortholog,并将其命名为dCATH。 dCATH的cDNA序列编码一个预测的146个氨基酸的多肽,该多肽由17个残基的信号肽,一个109个残基的保守的cathelin域和一个20个残基的成熟肽组成。系统发育分析表明,dCATH与其他禽类肽高度不同。该肽的α-螺旋结构在体外对多种细菌均表现出强大的抗菌活性,其最低抑菌浓度最低为2-4μM。此外,dCATH还显示出细胞毒性,在存在或不存在浓度为32μm或20μm的10%胎牛血清的情况下,裂解50%的哺乳动物红细胞,并以10μm的浓度杀死50%的HaCaT细胞。通过扫描电子显微镜和透射电子显微镜对细菌外膜和内膜的影响表明,dCATH通过增加渗透性杀死微生物细胞,从而导致膜完整性的丧失。

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