Identification and characterization of a hepcidin from half-smooth tongue sole Cynoglossus semilaevis.
Identification and characterization of a hepcidin from half-smooth tongue sole Cynoglossus semilaevis.
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DOI：
10.1016/j.fsi.2012.04.011
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发表时间：
2012-08
影响因子：
4.7
通讯作者：
Yanan Wang;Xudong Liu;Liman Ma;Yan Yu;Haiyang Yu;Shafi Mohammed;Guannan Chu;Linlin Mu;Quanqi Zhang
中科院分区：
文献类型：
--
作者：
Yanan Wang;Xudong Liu;Liman Ma;Yan Yu;Haiyang Yu;Shafi Mohammed;Guannan Chu;Linlin Mu;Quanqi Zhang
Hepcidin 是一种抗菌肽，具有先天免疫和铁调节的双重功能。在此，基于EST数据库的序列，我们从半滑舌鳎（Cynoglossus semilaevis）中分离并鉴定了一个hepcidin基因（简称CsHepcidin）。编码区分析表明CsHepcidin基因包含3个外显子和2个内含子。推定的 CsHepcidin 与其他 hepcidin 直系同源物表现出极大的相似性，特别是在其 24 个氨基酸信号肽、典型的 RX(K/R)R 基序和成熟阳离子肽中的 8 个保守半胱氨酸残基方面。系统发育分析表明CsHepcidin是棘翅目Hepcidin 1型肽，与Solea senegalensis hepcidin高度同源。在半滑鼠个体发育过程中，CsHepcidin mRNA在未受精卵中检测到较低水平，在孵化后6天增加，并在变态阶段显着下降。 CsHepcidin 转录物在大多数组织中显示出组成型基础表达，尤其是在肝脏中。用福尔马林灭活的鳗弧菌进行攻击导致肝脏、头肾和脾脏中 CsHepcidin 基因以时间依赖性方式显着上调。生物活性分析表明，重组 CsHEP 在体外对细菌病原体表现出直接的抗菌活性，特别是对主要鱼类病原体 V. anguillarum 和 Edwardsiella tarda 显示出很强的活性。所有这些结果表明，CsHepcidin 可能参与对微生物病原体入侵的初始反应。需要进一步探索以阐明 CsHepcidin 在半滑鼠铁调节和胚胎发生中的作用。
Hepcidin, an antimicrobial peptide, has a dual function including innate immunity and iron regulation. Here, based on the sequence of an EST database, we have isolated and characterized a hepcidin gene (referred to as CsHepcidin) from half-smooth tongue sole (Cynoglossus semilaevis). Analysis of the coding regions indicated CsHepcidin gene comprised 3 exons and 2 introns. The putative CsHepcidin showed a great similarity to other hepcidin orthologues, particularly with respect to its 24 aa signal peptide, typical RX(K/R)R motif and eight conserved cysteine residues in the mature cationic peptide. Phylogenic analysis indicated that CsHepcidin was a hepcidin 1-type peptide of acanthopterygians, with highly homologous with Solea senegalensis hepcidin. In C. semilaevis ontogeny, CsHepcidin mRNA was detected at a low level in unfertilized eggs, increased on 6 d after hatching, and decreased remarkably at metamorphic stage. CsHepcidin transcripts showed a constitutive basal expression in most of the tissues, especially in liver. Challenge with formalin-inactivated Vibrio anguillarum led to significantly up-regulations of CsHepcidin gene in liver, head kidney and spleen in time-dependent manners. Biological activity analysis showed that recombinant CsHEP exhibited direct antimicrobial activity against bacterial pathogens in vitro, particularly showed strong activity against the principal fish pathogens, V. anguillarum and Edwardsiella tarda. All these results suggest that CsHepcidin may be involved in the initial response to invasion of microbial pathogens. Further exploration to elucidate the role of CsHepcidin in iron regulation and embryogenesis in C. semilaevis are needed.