The role of serine- and metalloproteases in Nasonia vitripennis venom in cell death related processes towards a Spodoptera frugiperda Sf21 cell line
The role of serine- and metalloproteases in Nasonia vitripennis venom in cell death related processes towards a Spodoptera frugiperda Sf21 cell line
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DOI：
10.1016/j.jinsphys.2013.05.004
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发表时间：
2013-08-01
影响因子：
2.2
通讯作者：
de Graaf, Dirk C.
中科院分区：
文献类型：
--
作者：
Formesyn, Ellen M.;Heyninck, Karen;de Graaf, Dirk C.
蛋白酶是玻璃螯虾外寄生蜂的主要毒液成分。利用蛋白酶抑制剂灭活一个或两个蛋白酶家族，研究了丝氨酸蛋白酶和金属蛋白酶两个蛋白酶家族在frugiperda （Spodoptera frugiperda, Sf21）细胞系中可能的细胞毒性功能。对贴壁细胞的活力测定表明，这两个蛋白酶家族都是玻璃螯虾毒液的主要细胞毒性化合物。然而，毒后24小时对悬浮细胞进行的活力测定和流式细胞术显示，金属蛋白酶的失活并没有提高细胞存活率。这些结果表明，这两个蛋白酶家族可能具有组织特异性功能。时间过程实验表明，玻璃螯虾毒液丝氨酸蛋白酶参与诱导Sf21细胞株凋亡。然而，其他毒液化合物也可能参与这一过程，当一种特定类型的细胞死亡被抑制时，不同的细胞死亡途径可能会接管。在其天然宿主的寄生过程中，这两个蛋白酶家族可能在免疫相关过程和组织破坏中发挥作用，从而使毒液分布。综上所述，本研究对玻璃翅螨毒液中丝氨酸和金属蛋白酶的功能提供了重要的认识。(c) 2013 Elsevier Ltd.版权所有。
Proteases are predominant venom components of the ectoparasitoid Nasonia vitripennis. Two protease families, serine proteases and metalloproteases were examined for their possible cytotoxic functions in the Spodoptera frugiperda (Sf21) cell line using protease inhibitors that inactivate one or both protease families. Viability assays on adherent cells indicated that both protease families are among the main cytotoxic compounds of N. vitripennis venom. However, viability assays and flow cytometry performed on suspension cells 24 h after envenomation revealed that inactivation of metalloproteases did not improve cell survival. These results indicate that both protease families may have tissue specific functions. Time course experiments indicate that serine proteases of N. vitripennis venom are responsible for inducing apoptosis in the Sf21 cell line. However, other venom compounds could also be involved in this process and different cell death pathways may take over when a specific type of cell death is inhibited. During parasitation of their natural hosts, both protease families may possibly function in immune related processes and tissue destruction, enabling venom distribution. Overall, this study provides important insights into the functions of serine and metalloproteases in the venom of N. vitripennis. (c) 2013 Elsevier Ltd. All rights reserved.