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Proteomic profile and protease activity in the skin mucus of greater amberjack (Seriola dumerili) infected with the ectoparasite Neobenedenia girellae — an immunological approach
Fish & Shellfish Immunology ( IF 4.7 ) Pub Date : 2021-01-11 , DOI: 10.1016/j.fsi.2021.01.001
Álvaro Fernández-Montero , Silvia Torrecillas , Daniel Montero , Felix Acosta , María-José Prieto-Álamo , Nieves Abril , Juan Jurado

Skin mucus is considered the first barrier against diseases in fish. The skin mucus protein profile of the greater amberjack (Seriola dumerili) and its changes due to experimental infection with Neobenedenia girellae were studied by combining 2-DE-MS/MS and gel-free LC-MS/MS proteomic approaches. The 2-DE results led to the identification of 69 and 55 proteins in noninfected and infected fish, respectively, and revealed that keratins were specifically cleaved in parasitized fish. Therefore, the skin mucus of the infected fish showed a higher protease activity due to, at least in part, an increase of metal-dependent protease and serine-type protease activities. Additionally, through a gel-free LC-MS/MS analysis, 1377 and 1251 different proteins were identified in the skin mucus of healthy and parasitized fish, respectively. The functional analysis of these proteins demonstrated a statistical overrepresentation of ribosomal proteins (a well-known source of antimicrobial peptides) in N. girellae-infected fish. In contrast, the components of membranes and protein transport GO categories were underrepresented after infection. Immune system process-related proteins constituted 2.5% of the total skin mucosal proteins. Among these skin mucosal proteins, 14 and 15 proteins exclusive to non-parasitized and parasitized fish were found, respectively, including specific serine-type proteases and metalloproteases in the parasitized fish. Moreover, the finding of tryptic peptides exclusive to some bacterial genera, obtained by gel-free LC-MS/MS, allowed us to construct a preliminary map of the microbiota living in the mucus of S. dumerili, with Pseudomonas and Paracoccus the most represented genera in both noninfected and infected fish.



中文翻译:

感染外寄生虫新本尼德尼亚鞭毛虫的较大greater(Seriola dumerili)皮肤黏液中的蛋白质组学特征和蛋白酶活性—免疫学方法

皮肤粘液被认为是抵抗鱼类疾病的第一道屏障。大mber(Seriola dumerili)的皮肤粘液蛋白谱及其由于实验性感染新刺线虫的变化。通过结合2-DE-MS / MS和无凝胶LC-MS / MS蛋白质组学方法进行研究。2-DE结果导致分别在未感染和感染的鱼类中鉴定出69和55种蛋白质,并揭示了在被寄生的鱼类中角蛋白被特异性裂解。因此,至少部分地由于金属依赖性蛋白酶和丝氨酸型蛋白酶活性的增加,被感染鱼的皮肤粘液显示出较高的蛋白酶活性。此外,通过无胶LC-MS / MS分析,在健康和被寄生鱼的皮肤粘液中分别鉴定出1377和1251种不同的蛋白质。这些蛋白质的功能分析表明,在长鼻猪笼草中核糖体蛋白质(一种众所周知的抗菌肽来源)的统计含量过高感染的鱼。相反,感染后膜的成分和蛋白质转运GO类别的代表性不足。与免疫系统过程相关的蛋白质占皮肤粘膜总蛋白质的2.5%。在这些皮肤粘膜蛋白中,分别发现了14种和15种非寄生虫和寄生虫鱼类专有的蛋白质,包括寄生虫鱼中的特定丝氨酸型蛋白酶和金属蛋白酶。此外,通过无凝胶LC-MS / MS获得的某些细菌属专有的胰蛋白酶消化肽的发现,使我们能够构建生活在链球菌黏液的微生物群的初步图谱,其中假单胞菌球菌最为典型未感染和感染鱼的属。

更新日期:2021-01-11
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