Functional characterization and gene expression profile of perforin-2 in starry flounder (Platichthys stellatus)
| Autor: | Mi Young Cho, Dong-Hee Cho, Kwang-Min Choi, Myoung Sug Kim, Min-Soo Joo, Hyun-Ja Han, Hye-Sung Choi, Do-Hyung Kim, Chan-Il Park, Seong Don Hwang |
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| Rok vydání: | 2020 |
| Předmět: |
0301 basic medicine
Fish Proteins Pore Forming Cytotoxic Proteins Flounder Aquatic Science medicine.disease_cause 03 medical and health sciences Fish Diseases Gene expression medicine Environmental Chemistry Animals Amino Acid Sequence Escherichia coli Peptide sequence Starry flounder Phylogeny MACPF biology Gene Expression Profiling 04 agricultural and veterinary sciences General Medicine biology.organism_classification Platichthys Molecular biology Immunity Innate Perciformes 030104 developmental biology Perforin Gene Expression Regulation 040102 fisheries biology.protein 0401 agriculture forestry and fisheries Complement membrane attack complex Sequence Alignment |
| Zdroj: | Fishshellfish immunology. 107 |
| ISSN: | 1095-9947 |
| Popis: | 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). |
| Databáze: | OpenAIRE |
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