Key Laboratory of South China Sea Fishery Resources Exploitation & Utilization, Ministry of Agriculture and Rural Affairs, South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, Guangdong, 510300, PR China; Guangdong Provincial Key Laboratory of Fishery Ecology and Environment, South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, 510300, China.
Key Laboratory of South China Sea Fishery Resources Exploitation & Utilization, Ministry of Agriculture and Rural Affairs, South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, Guangdong, 510300, PR China; Guangdong Provincial Key Laboratory of Fishery Ecology and Environment, South China Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou, 510300, China.
Fish Shellfish Immunol. 2023 Apr;135:108674. doi: 10.1016/j.fsi.2023.108674. Epub 2023 Mar 16.
Cytochrome P450 (CYPs) enzymes are one of the critical detoxification enzymes, playing a key role in antioxidant defense. However, the information of CYPs cDNA sequences and their functions are lacked in crustaceans. In this study, a novel full-length of CYP2 from the mud crab (designated as Sp-CYP2) was cloned and characterized. The coding sequence of Sp-CYP2 was 1479 bp in length and encoded a protein containing 492 amino acids. The amino acid sequence of Sp-CYP2 comprised a conserved heme binding site and chemical substrate binding site. Quantitative real-time PCR analysis revealed that Sp-CYP2 was ubiquitously expressed in various tissues, and it was highest in the heart followed by the hepatopancreas. Subcellular localization showed that Sp-CYP2 was prominently located in the cytoplasm and nucleus. The expression of Sp-CYP2 was induced by Vibrio parahaemolyticus infection and ammonia exposure. During ammonia exposure, ammonia exposure can induce oxidative stress and cause severely tissue damage. Knocking down Sp-CYP2 in vivo can increase malondialdehyde content and the mortality of mud crabs after ammonia exposure. All these results suggested that Sp-CYP2 played a crucial role in the defense against environmental stress and pathogen infection in crustaceans.
细胞色素 P450（CYPs）酶是一种重要的解毒酶，在抗氧化防御中起着关键作用。然而，甲壳类动物中 CYPs cDNA 序列及其功能的信息仍然缺乏。在本研究中，我们克隆并鉴定了一种来自泥蟹的新型全长 CYP2（命名为 Sp-CYP2）。Sp-CYP2 的编码序列长 1479bp，编码一个含有 492 个氨基酸的蛋白质。Sp-CYP2 的氨基酸序列包含一个保守的血红素结合位点和化学底物结合位点。实时定量 PCR 分析显示 Sp-CYP2 在各种组织中广泛表达，在心组织中表达量最高，其次是肝胰腺。亚细胞定位显示 Sp-CYP2 主要位于细胞质和细胞核中。Sp-CYP2 的表达受副溶血弧菌感染和氨暴露诱导。在氨暴露期间，氨暴露会诱导氧化应激并导致组织严重损伤。体内敲低 Sp-CYP2 会增加氨暴露后泥蟹的丙二醛含量和死亡率。这些结果表明 Sp-CYP2 在甲壳类动物抵御环境胁迫和病原体感染方面发挥着关键作用。
