YPTB3816 of Yersinia pseudotuberculosis strain IP32953 is a virulence-related metallo-oligopeptidase.
YPTB3816 of Yersinia pseudotuberculosis strain IP32953 is a virulence-related metallo-oligopeptidase.
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DOI：
10.1186/s12866-016-0900-7
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发表时间：
2016-11-25
期刊：
影响因子：
4.2
通讯作者：
Karlyshev AV
中科院分区：
文献类型：
--
作者：
Atas A;Seddon AM;Ford DC;Cooper IA;Wren BW;Oyston PC;Karlyshev AV
尽管已知细菌肽酶是由包括病原菌在内的多种微生物产生的，但它们在细菌生理学中的作用尚不完全清楚。特别地，寡肽酶被认为主要参与短肽的降解，例如短肽。在经典蛋白质分泌途径中释放的前导肽。本研究的目的是研究假结核耶尔森氏菌寡肽酶编码基因 opdA 基因失活对体内和体外细菌特性的影响，并测试各个纯化酶的酶活性对不同二价阳离子存在的依赖性。在这项研究中，我们发现假结核耶尔森氏菌的寡肽酶 OpdA 是细菌毒力所必需的，而敲除相应基因对细菌体外活力或生长率没有任何影响。此外，我们研究了这种酶在大肠杆菌中表达和纯化后的酶学特性。使用去除污染物二价阳离子和不同类型的荧光标记底物的酶，我们发现其活性对特定阳离子的存在有很强的依赖性。出乎意料的是，Zn2+仅在低浓度下表现出刺激活性，但在较高浓度下则抑制酶。相比之下，Co2+、Ca2+ 和 Mn2+ 在所有测试浓度下均刺激活性，而 Mg2+ 显示在所有使用浓度下对酶活性均无影响。这项研究的结果为细菌肽酶的研究，特别是金属寡肽酶的研究提供了有价值的贡献。这是第一项证明假结核耶尔森氏菌的致病性需要 opdA 的研究。报告的数据对于更好地了解 OpdA 样酶在细菌感染发病机制中的作用非常重要。该蛋白质的表征可以作为开发基于该肽酶活性的特异性抑制的新型抗菌药物的基础。本文的在线版本 (doi:10.1186/s12866-016-0900-7) 包含补充材料，可供授权用户使用。
Although bacterial peptidases are known to be produced by various microorganisms, including pathogenic bacteria, their role in bacterial physiology is not fully understood. In particular, oligopeptidases are thought to be mainly involved in degradation of short peptides e.g. leader peptides released during classical protein secretion pathways. The aim of this study was to investigate effects of inactivation of an oligopeptidase encoding gene opdA gene of Yersinia pseudotuberculosis on bacterial properties in vivo and in vitro, and to test dependence of the enzymatic activity of the respective purified enzyme on the presence of different divalent cations. In this study we found that oligopeptidase OpdA of Yersinia pseudotuberculosis is required for bacterial virulence, whilst knocking out the respective gene did not have any effect on bacterial viability or growth rate in vitro. In addition, we studied enzymatic properties of this enzyme after expression and purification from E. coli. Using an enzyme depleted of contaminant divalent cations and different types of fluorescently labelled substrates, we found strong dependence of its activity on the presence of particular cations. Unexpectedly, Zn2+ showed stimulatory activity only at low concentrations, but inhibited the enzyme at higher concentrations. In contrast, Co2+, Ca2+ and Mn2+ stimulated activity at all concentrations tested, whilst Mg2+ revealed no effect on the enzyme activity at all concentrations used. The results of this study provide valuable contribution to the investigation of bacterial peptidases in general, and that of metallo-oligopeptidases in particular. This is the first study demonstrating that opdA in Yersinia pseudotuberculsosis is required for pathogenicity. The data reported are important for better understanding of the role of OpdA-like enzymes in pathogenesis in bacterial infections. Characterisation of this protein may serve as a basis for the development of novel antibacterials based on specific inhibition of this peptidase activity. The online version of this article (doi:10.1186/s12866-016-0900-7) contains supplementary material, which is available to authorized users.
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影响因子：
3.2
作者：
CONLIN, CA;MILLER, CG
通讯作者：
MILLER, CG
影响因子：
6.1
作者：
Kowalska, K;Carr, DB;Lipkowski, AW
通讯作者：
Lipkowski, AW
影响因子：
3.9
作者：
Lorenzon, Ricardo Z.;Cunha, Carlos E. L.;Carmona, Adriana K.
通讯作者：
Carmona, Adriana K.
影响因子：
3.2
作者：
Conlin, CA;Miller, CG
通讯作者：
Miller, CG
影响因子：
7
作者：
Olsen, JV;Ong, SE;Mann, M
通讯作者：
Mann, M