期刊
DEVELOPMENTAL AND COMPARATIVE IMMUNOLOGY
卷 117, 期 -, 页码 -出版社
ELSEVIER SCI LTD
DOI: 10.1016/j.dci.2020.103980
关键词
Procambarus clarkii; Masquerade; Trypsin-like serine protease; Prophenoloxidase; Antibacterial activity
资金
- National Natural Science Foundation of China [31902411]
- China Postdoctoral Science Foundation [2019M661955, 2020T130560]
- Jiangsu Province Postdoctoral Science Foundation [2019K189]
The study revealed that the secreted trypsin-like serine protease Mas in crayfish can enhance the immune response by binding with bacteria and activating the prophenoloxidase system. Interference with the expression of PcMas led to reduced phenoloxidase activity and increased mortality caused by A. hydrophila infection. Injection of recombinant protein of PcMas resulted in enhanced phenoloxidase activity and reduced mortality due to A. hydrophila infections.
Masquerade (Mas) is a secreted trypsin-like serine protease (SPs) and involved in immune response in some arthropods. However, according to previous studies, Mas presents different functional activities. In the present study, the functional mechanisms of Mas in crayfish Procambarus clarkii immune defense were studied. A fragment cDNA sequence of PcMas was identified and characterized. From the structural analysis, it contains a trypsin-like serine protease domain. The highest expression level of PcMas was detected in hepatopancreas. The infection of A. hydrophila could induce the expression of PcMas, while the WSSV infection did not cause changes in the expression of PcMas. Through the prokaryotic expression system, the PcMas protein was expressed in E. coli. It was verified that PcMas can bind to bacteria in vitro and inhibit the growth of the bacteria. By dsRNA interference with the expression of PcMas, the decrease expression of PcMas led to a decrease in the activity of phenoloxidase in hemolymph and an increase of mortality caused by A. hydrophila infection. The injection of recombinant protein can enhance the activity of phenoloxidase and reduce mortality caused by A. hydrophila infections. Therefore, the present study confirmed that PcMas could improve the body's immune response to eliminate bacterial pathogens by binding with bacteria and activating the prophenoloxidase system. The results will enrich the molecular mechanisms of crustaceans immune defense.
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