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银鲳(Pampus argenteus)中半胱氨酸蛋白酶抑制剂 M 的鉴定、分子特征分析及免疫功能研究。

Identification, molecular profiling and immune functions of cystatin M in silver pomfret (Pampus argenteus).

机构信息

Key Laboratory of Aquacultural Biotechnology (Ningbo University), Ministry of Education, Ningbo, Zhejiang, China.

College of Life Sciences, Yantai University, Yantai, China.

出版信息

Fish Shellfish Immunol. 2024 Oct;153:109844. doi: 10.1016/j.fsi.2024.109844. Epub 2024 Aug 20.

DOI:10.1016/j.fsi.2024.109844
PMID:39168290
Abstract

Cystatins play an important role in various physiological and pathological processes of organisms, including regulating protein metabolism, antigen processing, inflammatory response, nutritional disorders, and controlling enzyme activity. However, research on immunity functions of fish cystatin M is limited. In this study, Pampus argenteus cystatin M (Pacystatin M) was identified and analyzed. Its amino acid sequence was highly conserved in teleosts, and included the conserved cystatin cysteine protease inhibitor motifs. Pacystatin M was highly expressed in the gill, spleen, and intestine, whereas the expression levels of liver and kidney were lower. Furthermore, Nocardia seriolae infection up-regulated the expression of Pacystatin M in the kidney, spleen and liver, with particularly significant expression observed in the liver on day 15 post-infection. Functional analysis indicated that the recombinant Pacystatin M showed increasing inhibitory activity against papain within a certain concentration range, suggesting that the inhibition was likely competitive. Additionally, Pacystatin M demonstrated the ability to inhibit bacterial growth and high thermal stability. These results suggested that Pacystatin M might be involved in the immune response to microbial invasion and provided new reference addressing disease issues in the large-scale farming of silver pomfret.

摘要

组织蛋白酶抑制剂在生物体的各种生理和病理过程中发挥着重要作用,包括调节蛋白质代谢、抗原处理、炎症反应、营养紊乱和控制酶活性。然而,关于鱼类组织蛋白酶 M 的免疫功能的研究还很有限。在本研究中,鉴定并分析了中国团头鲂组织蛋白酶 M(Pacystatin M)。其氨基酸序列在硬骨鱼类中高度保守,包含保守的组织蛋白酶抑制剂结构域。Pacystatin M 在鳃、脾脏和肠道中高表达,而在肝脏和肾脏中的表达水平较低。此外,鮰爱德华氏菌感染可上调肾脏、脾脏和肝脏中 Pacystatin M 的表达,感染后第 15 天肝脏中的表达水平尤其显著。功能分析表明,重组 Pacystatin M 在一定浓度范围内对木瓜蛋白酶的抑制活性呈增加趋势,提示这种抑制可能是竞争性的。此外,Pacystatin M 具有抑制细菌生长和高热稳定性的能力。这些结果表明,Pacystatin M 可能参与了对微生物入侵的免疫反应,为大规模养殖银鲳的疾病问题提供了新的参考依据。

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