Department of Biochemistry, Faculty of Biology and Biotechnology, University of Warmia and Mazury in Olsztyn, 10-719 Olsztyn, Poland.
Clinical Pharmacology Group, Health Research Institute of Santiago de Compostela (IDIS), Santiago de Compostela, 15-706 A Coruña, Spain.
Int J Mol Sci. 2022 Apr 14;23(8):4336. doi: 10.3390/ijms23084336.
s. s. is a parasitic nematode of marine mammals and causative agent of anisakiasis in humans. The cuticle and intestine of the larvae are the tissues most responsible for direct and indirect contact, respectively, of the parasite with the host. At the L4 larval stage, tissues, such as the cuticle and intestine, are fully developed and functional, in contrast to the L3 stage. As such, this work provides for the first time the tissue-specific proteome of s. s. larvae in the L4 stage. Statistical analysis (FC ≥ 2; -value ≤ 0.01) showed that 107 proteins were differentially regulated (DRPs) between the cuticle and the rest of the larval body. In the comparison between the intestine and the rest of the larval body at the L4 stage, 123 proteins were identified as DRPs. Comparison of the individual tissues examined revealed a total of 272 DRPs, with 133 proteins more abundant in the cuticle and 139 proteins more abundant in the intestine. Detailed functional analysis of the identified proteins was performed using bioinformatics tools. Glycolysis and the tricarboxylic acid cycle were the most enriched metabolic pathways by cuticular and intestinal proteins, respectively, in the L4 stage of s. s. The presence of two proteins, folliculin (FLCN) and oxoglutarate dehydrogenase (OGDH), was confirmed by Western blot, and their tertiary structure was predicted and compared with other species. In addition, host-pathogen interactions were identified, and potential new allergens were predicted. The result of this manuscript shows the largest number of protein identifications to our knowledge using proteomics tools for different tissues of L4 larvae of s. s. The identified tissue-specific proteins could serve as targets for new drugs against anisakiasis.
旋毛线虫是一种海洋哺乳动物的寄生线虫，也是人类异尖线虫病的病原体。幼虫的角质层和肠道分别是寄生虫与宿主直接和间接接触的主要组织。在 L4 幼虫阶段，组织（如角质层和肠道）已完全发育和功能齐全，与 L3 阶段形成对比。因此，这项工作首次提供了 L4 阶段旋毛线虫幼虫组织特异性蛋白质组。统计分析（FC≥2；-值≤0.01）表明，角质层和幼虫其他部位之间有 107 种蛋白质差异表达（DRPs）。在 L4 阶段比较肠道和幼虫其他部位时，鉴定出 123 种 DRPs。比较单独检查的组织，共发现 272 种 DRPs，其中 133 种蛋白质在角质层中更为丰富，139 种蛋白质在肠道中更为丰富。使用生物信息学工具对鉴定出的蛋白质进行了详细的功能分析。糖酵解和三羧酸循环分别是 L4 期旋毛线虫角质层和肠道蛋白最丰富的代谢途径。Western blot 证实了两种蛋白质，即多囊泡蛋白（FLCN）和氧化戊二酸脱氢酶（OGDH）的存在，并对其三级结构进行了预测，并与其他物种进行了比较。此外，还鉴定了宿主-病原体相互作用，并预测了潜在的新过敏原。本文的结果显示，使用蛋白质组学工具对旋毛线虫 L4 幼虫的不同组织进行鉴定，是迄今为止鉴定到的蛋白质数量最多的一次。鉴定出的组织特异性蛋白质可以作为抗异尖线虫病新药的靶标。
