CLIPB10 is a Terminal Protease in the Regulatory Network That Controls Melanization in the African Malaria Mosquito Anopheles gambiae.
CLIPB10 is a Terminal Protease in the Regulatory Network That Controls Melanization in the African Malaria Mosquito Anopheles gambiae.
复制标题
DOI：
10.3389/fcimb.2020.585986
复制
发表时间：
2020
影响因子：
5.7
通讯作者：
Michel K
中科院分区：
文献类型：
--
作者：
Zhang X;Li M;El Moussawi L;Saab S;Zhang S;Osta MA;Michel K
动物的体液免疫反应通常受到调节蛋白分解的严格控制。这种蛋白分解是由胞外蛋白酶级联作用的，其激活的最终结果是关键免疫蛋白和酶的蛋白水解性切割。这种免疫系统调节的一个模型是昆虫的黑化反应，在这种反应中，原酚氧化酶(ProPO)的激活导致异物(如寄生虫、细菌和真菌)表面迅速形成真黑素。ProPO的激活受到一个所谓的Clip结构域丝氨酸蛋白酶网络的严格调控，这些酶是它们的蛋白水解性不活跃的同系物，以及它们的丝氨酸抑制剂。在撒哈拉以南非洲的主要疟疾媒介冈比亚按蚊中，对这一蛋白酶网络的操纵会影响对多种微生物的抗药性，以及宿主的生存。然而，到目前为止，我们对蚊子中蛋白水解酶网络的分子组成和调控的了解是有限的。在这里，我们报告了CLIP结构域丝氨酸蛋白酶CLIPB10在这个网络中的功能，结合了遗传和生化分析。CLIPB10基因敲除可在无感染的情况下部分逆转Serpin 2沉默诱导的黑色素瘤形成。在难治的蚊子遗传背景下，CLIPB10也是啮齿动物疟疾寄生虫伯氏疟原虫运动阶段黑化所必需的部分。重组丝裂蛋白2蛋白，一个关键的抑制子，在前激活级联在AN。在体外，重组活化的CLIPB10能有效地抑制CLIPB10的活性，其化学计量比为1.89：1。重组活化的CLIPB10能在体外提高六爪虾血淋巴中PO的活性，并能在体外直接激活纯化后的六分枝杆菌。综上所述，这些数据证实CLIPB10是AN中第二个具有酚氧化酶原激活功能的蛋白酶。冈比亚，除了先前描述的CLIPB9外，还表明控制黑化的蛋白酶网络中存在功能冗余。此外，我们的数据表明，寄生虫的组织黑化和体液黑化至少部分是由相同的蛋白酶介导的。
Humoral immune responses in animals are often tightly controlled by regulated proteolysis. This proteolysis is exerted by extracellular protease cascades, whose activation culminates in the proteolytic cleavage of key immune proteins and enzymes. A model for such immune system regulation is the melanization reaction in insects, where the activation of prophenoxidase (proPO) leads to the rapid formation of eumelanin on the surface of foreign entities such as parasites, bacteria and fungi. ProPO activation is tightly regulated by a network of so-called clip domain serine proteases, their proteolytically inactive homologs, and their serpin inhibitors. In Anopheles gambiae, the major malaria vector in sub-Saharan Africa, manipulation of this protease network affects resistance to a wide range of microorganisms, as well as host survival. However, thus far, our understanding of the molecular make-up and regulation of the protease network in mosquitoes is limited. Here, we report the function of the clip domain serine protease CLIPB10 in this network, using a combination of genetic and biochemical assays. CLIPB10 knockdown partially reversed melanotic tumor formation induced by Serpin 2 silencing in the absence of infection. CLIPB10 was also partially required for the melanization of ookinete stages of the rodent malaria parasite Plasmodium berghei in a refractory mosquito genetic background. Recombinant serpin 2 protein, a key inhibitor of the proPO activation cascade in An. gambiae, formed a SDS-stable protein complex with activated recombinant CLIPB10, and efficiently inhibited CLIPB10 activity in vitro at a stoichiometry of 1.89:1. Recombinant activated CLIPB10 increased PO activity in Manduca sexta hemolymph ex vivo, and directly activated purified M. sexta proPO in vitro. Taken together, these data identify CLIPB10 as the second protease with prophenoloxidase-activating function in An. gambiae, in addition to the previously described CLIPB9, suggesting functional redundancy in the protease network that controls melanization. In addition, our data suggest that tissue melanization and humoral melanization of parasites are at least partially mediated by the same proteases.
登录
查看更多内容
影响因子：
6.7
作者：
Habtewold T;Povelones M;Blagborough AM;Christophides GK
通讯作者：
Christophides GK
影响因子：
32.4
作者：
Frolet, Cecile;Thoma, Martine;Levashina, Elena A.
通讯作者：
Levashina, Elena A.
影响因子：
3.6
作者：
Hillyer, JF;Schmidt, SL;Christensen, BM
通讯作者：
Christensen, BM
影响因子：
56.9
作者：
Christophides, GK;Zdobnov, E;Kafatos, FC
通讯作者：
Kafatos, FC
影响因子：
4.8
作者：
Kan, Hongnan;Kim, Chan-Hee;Lee, Bok Luel
通讯作者：
Lee, Bok Luel