Extremely Abundant Antimicrobial Peptides Existed in the Skins of Nine Kinds of Chinese Odorous Frogs
Extremely Abundant Antimicrobial Peptides Existed in the Skins of Nine Kinds of Chinese Odorous Frogs
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九种中华臭蛙皮肤中含有极其丰富的抗菌肽
DOI：
10.1021/pr200782u
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发表时间：
2012-01-01
影响因子：
4.4
通讯作者：
Zhang, Yun
中科院分区：
文献类型：
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作者：
Yang, Xinwang;Lee, Wen-Hui;Zhang, Yun
由于其独特的作用机制，肽类药物被视为解决致病性微生物对经典抗生素产生危及生命的耐药性的有希望的候选药物。对两栖动物皮肤分泌物中天然抗菌肽（AMPs）的肽组学和基因组学研究可为设计具有治疗潜力的肽类抗生素提供大量结构 - 功能信息。在本研究中，我们从9种中国臭蛙的皮肤中鉴定出了大量抗菌肽。从3种不同的臭蛙中纯化出80种抗菌肽，并通过肽组学分析得到确认。我们的研究结果表明，抗菌肽的翻译后修饰在臭蛙中很少发生。从构建的9种中国臭蛙抗菌肽cDNA文库中克隆出了编码728种抗菌肽前体的cDNA，包括已确认的80种天然肽的所有前体。根据推导的成熟肽的序列相似性，这728种抗菌肽被分为97个不同的家族，其中鉴定出71个新家族。在这728种抗菌肽中，662种是新发现的，28种先前在其他蛙类物种中已有报道。我们的研究结果显示，相同的抗菌肽在臭蛙中广泛分布；目前鉴定出的49种抗菌肽可在不同的两栖动物物种中找到相同的分子。纯化的肽对4种受试微生物菌株显示出强大的抗菌活性。合成了23种推导的肽，并对其生物活性，包括抗菌、抗氧化、溶血、免疫调节和胰岛素释放活性进行了评估。我们的研究结果证明了两栖动物皮肤中抗菌肽的极度多样性，并为开发新型肽类抗生素提供了大量模板。
Peptide agents are regarded as hopeful candidates to solve life-threatening resistance of pathogenic microorganisms to classic antibiotics due to their unique action mechanisms. Peptidomic and genomic investigation of natural antimicrobial peptides (AMPs) from amphibian skin secretions can provide a large amount of structure-functional information to design peptide antibiotics with therapeutic potential. In the present study, we identified a large number of AMPs from the skins of nine kinds of Chinese odorous frogs. Eighty AMPs were purified from three different odorous frogs and confirmed by peptidomic analysis. Our results indicated that post-translational modification of AMPs rarely happened in odorous frogs. cDNAs encoding precursors of 728 AMPs, including all the precursors of the confirmed 80 native peptides, were cloned from the constructed AMP cDNA libraries of nine Chinese odorous frogs. On the basis of the sequence similarity of deduced mature peptides, these 728 AMPs were grouped into 97 different families in which 71 novel families were identified. Out of these 728 AMPs, 662 AMPs were novel and 28 AMPs were reported previously in other frog species. Our results revealed that identical AMPs were widely distributed in odorous frogs; 49 presently identified AMPs could find their identical molecules in different amphibian species. Purified peptides showed strong antimicrobial activities against 4 tested microbe strains. Twenty-three deduced peptides were synthesized and their bioactivities, including antimicrobial, antioxidant, hemolytic, immunomodulatory and insulin-releasing activities, were evaluated. Our findings demonstrate the extreme diversity of AMPs in amphibian skins and provide plenty of templates to develop novel peptide antibiotics,