Department of Marine Biology & Aquaculture, College of Marine Science, Gyeongsang National University, 455, Tongyeong, 650-160, Republic of Korea.
Pathology Research Division, National Institute of Fisheries Science, 408-1 Sirang-ri, Gijang-up, Gijang-gun, Busan, 46083, Republic of Korea.
Fish Shellfish Immunol. 2020 Dec;107(Pt B):511-518. doi: 10.1016/j.fsi.2020.11.011. Epub 2020 Nov 17.
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).
膜攻击复合物/穿孔素 (MACPF) 超家族由多功能蛋白组成，这些蛋白在微生物的膜表面形成孔，诱导其死亡，并具有各种免疫相关功能。PFN2 是一种具有 MACPF 结构域的穿孔素样蛋白，人类 PFN2 水平不足会增加对细菌感染的易感性，这可能对一些患者造成致命后果。因此，在本研究中，我们证实了 PFN2 在星斑川鲽（Platichthys stellatus）中的抗菌功能。根据已验证的 PsPFN2 氨基酸序列确认了分子特性。此外，还证实了组织特异性和病原体特异性 PsPFN2 mRNA 的表达特征。使用大肠杆菌生产重组蛋白，并确认其抗菌活性。PFN2 的编码序列（PsPFN2）在星斑川鲽中由 710 个残基组成。通过多重序列比对和系统发育分析表明，MACPF 结构域在进化过程中是保守的。在健康星斑川鲽的免疫相关器官（如脾脏和鳃）中大量分布 PsPFN2 mRNA，在人工感染细菌或病毒后证实其表达发生显著变化。我们克隆了 PFN2 的 MACPF 结构域区域，以产生重组蛋白（rPFN2），并证实其对多种细菌物种和寄生虫（Miamiensis avidus）的抗菌作用。
