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Functional characterization and gene expression profile of perforin-2 in starry flounder (Platichthys stellatus)
Fish & Shellfish Immunology ( IF 4.7 ) Pub Date : 2020-11-17 , DOI: 10.1016/j.fsi.2020.11.011
Kwang-Min Choi , Dong-Hee Cho , Min-Soo Joo , Hye-Sung Choi , Myoung Sug Kim , Hyun-Ja Han , Mi Young Cho , Seong Don Hwang , Do-Hyung Kim , Chan-Il Park

The membrane attack complex/perforin (MACPF) superfamily consists of multifunctional proteins that form pores on the membrane surface of microorganisms to induce their death and have various immune-related functions. PFN2 is a perforin-like protein with an MACPF domain, and humans with deficient PFN2 levels have increased susceptibility to bacterial infection, which can lead to fatal consequences for some patients. Therefore, in this study, we confirmed the antimicrobial function of PFN2 in starry flounder (Platichthys stellatus). The molecular properties were confirmed based on the verified amino acid sequence of PsPFN2. In addition, the expression characteristics of tissue-specific and pathogen-specific PsPFN2 mRNA were also confirmed. The recombinant protein was produced using Escherichia coli, and the antimicrobial activity was then confirmed. The coding sequence of PFN2 (PsPFN2) in P. stellatus consists of 710 residues. The MACPF domain was conserved throughout evolution, as shown by multiple sequence alignment and phylogenetic analysis. PsPFN2 mRNA is abundantly distributed in immune-related organs such as the spleen and gills of healthy starry flounder, and significant expression changes were confirmed after artificial infection by bacteria or viruses. We cloned the MACPF domain region of PFN2 to produce a recombinant protein (rPFN2) and confirmed its antibacterial effect against a wide range of bacterial species and the parasite (Miamiensis avidus).



中文翻译:

Perforin-2在繁星比目鱼(Platichthys stellatus)中的功能表征和基因表达谱

膜攻击复合物/穿孔素(MACPF)超家族由多功能蛋白质组成,这些蛋白质在微生物的膜表面形成孔以诱导其死亡并具有多种免疫相关功能。PFN2是具有MACPF结构域的穿孔素样蛋白,PFN2水平不足的人对细菌感染的敏感性增加,可能对某些患者造成致命后果。因此,在这项研究中,我们证实了PFN2在繁星比目鱼(Platichthys stellatus)中的抗菌功能。通过验证的PsPFN2氨基酸序列证实了分子性质。另外,组织特异性和病原体特异性的表达特征还证实了PsPFN2 mRNA。用大肠杆菌生产重组蛋白,然后确认其抗菌活性。的编码序列PFN2PsPFN2)在P.鲽由710个残基。MACPF结构域在整个进化过程中都是保守的,如多重序列比对和系统发育分析所示。PsPFN2 mRNA大量分布于免疫相关器官,例如健康星空比目鱼的脾脏和腮中,经细菌或病毒人工感染后,证实了明显的表达变化。我们克隆的MACPF域区域PFN2生产的重组蛋白(rPFN2),并确认了其对多种细菌和寄生虫(Miamiensis avidus)的抗菌作用。

更新日期:2020-11-19
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