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Dephosphorylation of T517 on Hemocyanin Is Required for Antibacterial Activity in Penaeus vannamei 
Qian Feng; Jude Juventus Aweya; Yue-Qian Huang; Pei Zhang; Fan Wang 
J Immunol (2023) 210 (9): 1396–1407.
https://doi.org/10.4049/jimmunol.2200598


Abstract
Posttranslational modifications expand the functions of immune-related proteins, especially during infections. The respiratory glycoprotein, hemocyanin, has been implicated in many other functions, but the role of phosphorylation modification in its functional diversity is not fully understood. In this study, we show that Penaeus vannamei hemocyanin (PvHMC) undergoes phosphorylation modification during bacterial infection. Dephosphorylation of PvHMC mediated by P. vannamei protein phosphatase 2A catalytic increases its in vitro antibacterial activity, whereas phosphorylation by P. vannamei casein kinase 2 catalytic subunit α decreases its oxygen-carrying capacity and attenuates its in vitro antibacterial activity. Mechanistically, we show that Thr517 is a critical phosphorylation modification site on PvHMC to modulate its functions, which when mutated attenuates the action of P. vannamei casein kinase 2 catalytic subunit α and P. vannamei protein phosphatase 2A catalytic, and hence abolishes the antibacterial activity of PvHMC. Our results reveal that phosphorylation of PvHMC modulates its antimicrobial functions in penaeid shrimp.


翻譯后修飾擴(kuò)大了免疫相關(guān)蛋白的功能，尤其是在感染期間。呼吸糖蛋白血藍(lán)蛋白與許多其他功能有關(guān)，但磷酸化修飾在其功能多樣性中的作用尚不完全清楚。在本研究中，我們發(fā)現(xiàn)南美白對蝦血藍(lán)蛋白（PvHMC）在細(xì)菌感染過程中發(fā)生磷酸化修飾。凡納濱對蝦蛋白磷酸酶2A催化介導(dǎo)的PvHMC的去磷酸化增加了其體外抗菌活性，而凡納濱濱對蝦酪蛋白激酶2催化亞基α的磷酸化降低了其載氧能力并減弱了其體外抑菌活性。從機(jī)制上講，我們發(fā)現(xiàn)Thr517是PvHMC上調(diào)節(jié)其功能的關(guān)鍵磷酸化修飾位點(diǎn)，當(dāng)突變時，它減弱了凡納濱對蝦酪蛋白激酶2催化亞基α和凡納濱濱對蝦蛋白磷酸酶2A催化的作用，從而消除了PvHMC的抗菌活性。我們的研究結(jié)果表明，PvHMC的磷酸化調(diào)節(jié)其在對蝦中的抗菌功能。