Identification and functional characterization of a novel ferritin subunit from the tropical sea cucumber, Stichopus monotuberculatus.
Identification and functional characterization of a novel ferritin subunit from the tropical sea cucumber, Stichopus monotuberculatus.
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DOI：
10.1016/j.fsi.2014.03.022
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发表时间：
2014-05
影响因子：
4.7
通讯作者：
C. Ren;Ting Chen;Xiao Jiang;Yanhong Wang;Chao-qun Hu
中科院分区：
文献类型：
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作者：
C. Ren;Ting Chen;Xiao Jiang;Yanhong Wang;Chao-qun Hu
铁蛋白是一种主要的无害铁储存蛋白，存在于大多数动物、植物和微生物的细胞类型中。本研究从海参中分离到一个铁蛋白亚基，命名为StmFer，并对其功能进行了鉴定。StmFer全长1184hp，5‘非翻译区131hp，3’非翻译区531hp，开放阅读框522hp，编码173个氨基酸的蛋白质，推测其相对分子质量为19.95 kDa。StmFer既具有脊椎动物铁蛋白重亚单位的铁氧合酶中心，又具有脊椎动物铁蛋白轻亚单位的铁成核部位。在基因结构上，StmFer只包含三个外显子，被两个内含子隔开。Northern印迹分析显示，肠组织和体腔细胞中均有较高水平的mRNA表达。在体腔细胞的体外实验中，StmFer的转录表达在攻击后3h对多聚核苷多核糖酸[Poly(I：C)]的反应最强(9.08倍)，其次是脂多糖(LPS)、氯化亚铁(FeCl2)和灭活细菌(溶藻弧菌)(分别为7.84、7.41和4.90倍)。此外，本研究还对重组铁蛋白的抗氧化活性和铁结合能力进行了研究。总体而言，我们的研究表明，海参铁蛋白不仅在细胞和组织铁的动态平衡中发挥关键作用，而且在天然免疫防御中也发挥着关键作用。
Ferritin is one of the major non-harm iron storage proteins that found in most cell types of animals, plants and microorganisms. In this study, a ferritin subunit named StmFer was identified from the sea cucumber (Stichopus monotuberculatus) and characterized functionally. The full-length cDNA of StmFer is 1184 bp in size with a 5′-untranslated region (UTR) of 131 bp, a 3′-UTR of 531 bp and an open reading frame of 522 bp that encoding a protein of 173 amino acids with a deduced molecular weight of 19.95 kDa. StmFer possesses both the ferroxidase center of vertebrate ferritin heavy subunit and iron nucleation sites of vertebrate ferritin light subunit. For the gene structure, StmFer contains only three exons separated by two introns. Higher levels of mRNA expression were noticed in intestine and coelomocyte ofS. monotuberculatusby northern blot analysis. Inin vitroexperiments performed in coelomocytes, transcriptional expression of StmFer showed the strongest response to polyriboinosinic polyribocytidylic acid [Poly (I:C)] (9.08 fold up-regulation), followed by lipopolysaccharides (LPS), ferrous chloride (FeCl2) and inactivated bacteria (Vibrio alginolyticus) (7.84, 7.41 and 4.90 fold up-regulation, respectively) after 3 h post-challenge. In addition, the anti-oxidation activity and iron binding capacity of recombinant ferritin protein were demonstrated in this study. As a whole, our study suggested that the ferritin from sea cucumber may play critical roles not only in the cellular and organismic iron homeostasis, but also in the innate immune defense.