Burkholderia cenocepacia ZmpB is a broad-specificity zinc metalloprotease involved in virulence
Burkholderia cenocepacia ZmpB is a broad-specificity zinc metalloprotease involved in virulence
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DOI：
10.1128/iai.00297-06
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发表时间：
2006-07-01
影响因子：
3.1
通讯作者：
Sokol, P. A.
中科院分区：
文献类型：
--
作者：
Kooi, C.;Subsin, B.;Sokol, P. A.
在以前的研究中，我们的特点伯克霍尔德菌cenocepacia ZmpA锌金属蛋白酶。在这项研究中，我们确定了B。cenocepacia具有另外的金属蛋白酶，我们将其命名为ZmpB。zmp B基因存在于与ZmpA相同的物种中，并在B中检测到。洋葱B. cepacia，B. cenocepacia，B. stabilis，B. ambifaria和B. pyrrocinia，但缺席B。multivorans，B. vietnamiensis，B. dolosa和B. anthina。表达zmpB基因，并通过使用pPROEXHTa His（6）Tag表达系统从大肠杆菌纯化ZmpB。ZmpB具有预测的嗜热菌蛋白酶样蛋白酶典型的前酶原结构，并且与蜡状芽孢杆菌杆菌溶素有远亲关系。ZmpB表达为63 kDa的前酶原前体，其被自催化裂解成成熟的ZmpB（35 kDa）和27 kDa的前原肽。EDTA、1，10-菲咯啉和Zn 2+阳离子抑制ZmpB酶活性，表明它是一种金属蛋白酶。ZmpB对α-1蛋白酶抑制剂、α 2-巨球蛋白、IV型胶原、纤连蛋白、乳铁蛋白、转铁蛋白和免疫球蛋白具有蛋白水解活性。B。新洋葱菌zmp B和zmpA zmp B突变体对酪蛋白没有蛋白水解活性，并且在大鼠琼脂珠慢性感染模型中毒性较低，表明zmp B参与B。新洋葱毒力zmpB的表达受到CepIR和CciIR群体感应系统的调节。
In previous studies we characterized the Burkholderia cenocepacia ZmpA zinc metalloprotease. In this study, we determined that B. cenocepacia has an additional metalloprotease, which we designated ZmpB. The zmpB gene is present in the same species as ZmpA and was detected in B. cepacia, B. cenocepacia, B. stabilis, B. ambifaria, and B. pyrrocinia but was absent from B. multivorans, B. vietnamiensis, B. dolosa, and B. anthina. The zmpB gene was expressed, and ZmpB was purified from Escherichia coli by using the pPROEXHTa His(6) Tag expression system. ZmpB has a predicted preproenzyme structure typical of thermolysin-like proteases and is distantly related to Bacillus cereus bacillolysin. ZmpB was expressed as a 63-kDa preproenzyme precursor that was autocatalytically cleaved into mature ZmpB (35 kDa) and a 27-kDa prepropeptide. EDTA, 1,10-phenanthroline, and Zn2+ cations inhibited ZmpB enzyme activity, indicating that it is a metalloprotease. ZmpB had proteolytic activity against alpha-1 proteinase inhibitor, alpha 2-macrogobulin, type IV collagen, fibronectin, lactoferrin, transferrin, and immunoglobulins. B. cenocepacia zmpB and zmpA zmpB mutants had no proteolytic activity against casein and were less virulent in a rat agar bead chronic infection model, indicating that zmpB is involved in B. cenocepacia virulence. Expression of zmpB was regulated by both the CepIR and CciIR quorum-sensing systems.