School of Marine Science and Engineering, Qingdao Agricultural University, Qingdao, 266109, People's Republic of China.
School of Fisheries, Aquaculture and Aquatic Sciences, Auburn University, Auburn, AL, 36849, USA.
Fish Shellfish Immunol. 2020 Feb;97:322-335. doi: 10.1016/j.fsi.2019.12.005. Epub 2019 Dec 2.
Cathepsin L (CTSL) is one of the crucial enzymes in cathepsin family, which has been widely known for its involvement in the innate immunity. However, it still remains poorly understood how CTSL modulates the immune system of teleosts. In this study, we captured three cathepsin L genes (SmCTSL, SmCTSL.1 and SmCTSL1) from turbot (Scophthalmus maximus). The coding sequences of SmCTSL, SmCTSL.1 and SmCTSL1 are 1,026 bp, 1,005 bp and 1,017 bp in length and encode 341, 334 and 338 amino acids, respectively. In details, transcripts of CTSL genes share same domains as other CTSL genes, one signal peptide, one propeptide and one papain family cysteine protease domain. Protein interaction network analysis indicated that turbot CTSL genes may play important roles in apoptotic signaling and involve in innate immune response. Evidence from subcellular localization demonstrated that the three Cathepsin L proteins were ubiquitous in nucleus and cytoplasm. The cathepsin L genes were widely expressed in all the tested tissues with the highest expression level of SmCTSL in spleen, and SmCTSL.1 and SmCTSL1 in intestine. Following Vibrio anguillarum, Edwardsiella tarda and Streptococcus iniae challenge, these cathepsin L genes were significantly regulated in mucosal tissues in all the challenges, especially significant down-regulation occurred rapidly in intestine in all the three challenges. In addition, the three cathepsin L genes showed strong binding ability to all the examined microbial ligands (LPS, PGN and LTA). Further studies should be used to analyze the specific function of these three cathepsin L genes. By then, we can use their function to maintain the integrity of the mucosal barrier, thereby promoting the disease resistance line and family selection in turbot.
组织蛋白酶 L(CTSL)是组织蛋白酶家族中的关键酶之一,其在天然免疫中的作用已被广泛认识。然而,CTSL 如何调节硬骨鱼类的免疫系统仍知之甚少。在这项研究中,我们从大菱鲆(Scophthalmus maximus)中捕获了三个组织蛋白酶 L 基因(SmCTSL、SmCTSL.1 和 SmCTSL1)。SmCTSL、SmCTSL.1 和 SmCTSL1 的编码序列分别为 1026bp、1005bp 和 1017bp,分别编码 341、334 和 338 个氨基酸。具体而言,CTSL 基因的转录本与其他 CTSL 基因具有相同的结构域,包括一个信号肽、一个前肽和一个木瓜蛋白酶家族半胱氨酸蛋白酶结构域。蛋白质相互作用网络分析表明,大菱鲆 CTSL 基因可能在细胞凋亡信号转导中发挥重要作用,并参与固有免疫反应。亚细胞定位的证据表明,这三种组织蛋白酶 L 蛋白在细胞核和细胞质中普遍存在。这些组织蛋白酶 L 基因在所有检测组织中广泛表达,SmCTSL 在脾脏中的表达水平最高,SmCTSL.1 和 SmCTSL1 在肠道中的表达水平最高。在受到鳗弧菌、迟缓爱德华菌和链球菌感染后,这些组织蛋白酶 L 基因在所有挑战中均在黏膜组织中受到显著调控,尤其是在所有三种挑战中,肠道中的下调非常迅速。此外,这三种组织蛋白酶 L 基因对所有检测到的微生物配体(LPS、PGN 和 LTA)均具有很强的结合能力。进一步的研究应用于分析这三个组织蛋白酶 L 基因的特定功能。届时,我们可以利用它们的功能来维持黏膜屏障的完整性,从而促进大菱鲆的抗病性和家族选择。