The sea cucumber Apostichopus japonicus is an economically important species owing to its high nutritive and medicinal value. In order to avoid the pollution resulting from the overuse of antibiotics in A. japonicus aquaculture, various immunostimulants have been used as an alternative to improve the efficiency of A. japonicus farming. Our previous proteomic investigation has shown that several proteins participating in the immune-related physiology of A. japonicus were differentially expressed in the intestinal tissue in response to tussah immunoreactive substances (TIS). This study further explored the immunostimulation mechanism of TIS in A. japonicus. Phosphoproteomics technology was used to investigate the effect of TIS on protein phosphorylation in the intestine of A. japonicus following feeding with a TIS-supplemented diet. A total of 213 unique phosphoproteins were detected from 225 unique phosphopeptides. KEGG pathway analysis showed that majority of the phosphoproteins are involved in endocytosis, carbon metabolism and spliceosome functional group. Sixteen of the phosphoproteins exhibited differential phosphorylation in response to TIS and 12 of these were found to associate with biological functions. Of these 12 phosphoproteins, eight exhibited enhanced phosphorylation while four displayed reduced phosphorylation. These 12 proteins were further analyzed and all were found to play a role in regulating some aspects of the immune system and the growth of sea cucumbers, especially in phagocytosis, energy metabolism and disease resistance. The findings of this study could therefore shed new light on the immune pathways of sea cucumber that are affected by TIS. This could help us to better understand the underlying mechanism linked to the immunoenhancement of A. japonicus in response to TIS, one that is associated with the change in protein phosphorylation.

海参刺参(Apostichopus japonicus )具有较高的营养和药用价值，是一种重要的经济品种。为了避免青蒿养殖过度使用抗生素造成的污染，各种免疫刺激剂被用作替代品，以提高青蒿养殖效率。我们之前的蛋白质组学研究表明，参与A. japonicus免疫相关生理学的几种蛋白质在肠道组织中差异表达，以响应柞蚕免疫反应物质 (TIS)。本研究进一步探讨了 TIS 对A. japonicus的免疫刺激机制. 应用磷酸蛋白质组学技术研究 TIS 对日本刺参肠道蛋白磷酸化的影响喂食添加 TIS 的饮食后。从 225 种独特的磷酸肽中总共检测到 213 种独特的磷蛋白。KEGG通路分析表明，大部分磷蛋白参与胞吞作用、碳代谢和剪接体功能组。16 种磷蛋白在 TIS 反应中表现出不同的磷酸化，其中 12 种被发现与生物功能相关。在这 12 种磷蛋白中，8 种表现出增强的磷酸化，而 4 种表现出磷酸化降低。进一步分析了这 12 种蛋白质，发现它们都在调节免疫系统和海参生长的某些方面发挥作用，特别是在吞噬作用、能量代谢和抗病能力方面。因此，这项研究的结果可以为受 TIS 影响的海参的免疫途径提供新的线索。这可以帮助我们更好地理解与免疫增强相关的潜在机制。A. japonicus对 TIS 的响应，一种与蛋白质磷酸化的变化有关。