Molecular characterization and innate immune defense functional analysis of tartrate-resistant acid phosphatase 5 in Trachinotus ovatus

IF 3.2 2区 农林科学 Q1 FISHERIES
Reng Qiu , Jinge Wang , Qiaoli Yang , Ying Wu , Chunsheng Liu , Yang Lei , Yun Sun
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Abstract

The enzyme tartrate-resistant acid phosphatase 5 (TRAP5) performs an important biological function in immune defense. In the present study, the TRAP5 of Trachinotus ovatus (golden pompano, TroTRAP5) was cloned to assess its biological function. The open reading frame of TroTRAP5 was 990 bp in length and its mature peptide encoded a 34.775-kDa protein that contained a metallophosphatase superfamily domain. Homology sequence alignment revealed that the similarity among teleost fish was 57.66 %–75.99 %. Meanwhile, phylogenetic tree analysis showed that TroTRAP5 was clustered into one branch with teleost TRAP5. TroTRAP5 expression was the most abundant in immune organs (liver, spleen, head kidney, and gills) and up-regulated during bacterial infection. Of note, purified recombinant TroTRAP5 (rTroTRAP5) exhibited phosphatase activity, and the optimal pH and temperature for its activity were 6.0 and 50 ℃. In vivo experiments indicated that TroTRAP5 overexpression in T. ovatus significantly reduced bacterial infection in fish tissues, whereas TroTRAP5 knockdown increased bacterial dissemination and colonization in fish tissues. In summary, the present study results suggest that TroTRAP5 participates in the antibacterial immune response.
卵棘鲷抗酒石酸磷酸酶 5 的分子特征和先天免疫防御功能分析
抗酒石酸磷酸酶 5(TRAP5)在免疫防御中发挥着重要的生物学功能。本研究克隆了金鲳的 TRAP5,以评估其生物学功能。TroTRAP5的开放阅读框长度为990 bp,其成熟肽编码一个34.775-kDa的蛋白,含有金属磷酸酶超家族结构域。同源序列比对结果表明,TroTRAP5在远缘鱼类中的相似度为57.66%-75.99%。同时,系统发生树分析表明,TroTRAP5与远缘鱼类的TRAP5聚为一个分支。TroTRAP5在免疫器官(肝脏、脾脏、头肾和鳃)中的表达量最高,并在细菌感染时上调。值得注意的是,纯化的重组 TroTRAP5(rTroTRAP5)具有磷酸酶活性,其活性的最佳 pH 值和温度分别为 6.0 和 50 ℃。体内实验表明,TroTRAP5 在卵形栉水母中的过表达能显著减少细菌在鱼体组织中的感染,而 TroTRAP5 的敲除则会增加细菌在鱼体组织中的传播和定殖。总之,本研究结果表明,TroTRAP5 参与了抗菌免疫反应。
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来源期刊
Aquaculture Reports
Aquaculture Reports Agricultural and Biological Sciences-Animal Science and Zoology
CiteScore
5.90
自引率
8.10%
发文量
469
审稿时长
77 days
期刊介绍: Aquaculture Reports will publish original research papers and reviews documenting outstanding science with a regional context and focus, answering the need for high quality information on novel species, systems and regions in emerging areas of aquaculture research and development, such as integrated multi-trophic aquaculture, urban aquaculture, ornamental, unfed aquaculture, offshore aquaculture and others. Papers having industry research as priority and encompassing product development research or current industry practice are encouraged.
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