Mycoplasma agalactiae MAG_5040 is a Mg2+-dependent, sugar-nonspecific SNase recognised by the host humoral response during natural infection.
Mycoplasma agalactiae MAG_5040 is a Mg2+-dependent, sugar-nonspecific SNase recognised by the host humoral response during natural infection.
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DOI：
10.1371/journal.pone.0057775
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Alberti A
中科院分区：
文献类型：
--
作者：
Cacciotto C;Addis MF;Coradduzza E;Carcangiu L;Nuvoli AM;Tore G;Dore GM;Pagnozzi D;Uzzau S;Chessa B;Pittau M;Alberti A
在这项研究中，对无乳支原体 MAG_5040（一种与葡萄球菌 SNase 相似的镁依赖性核酸酶同源物）的酶活性进行了表征，并确定了其在自然感染期间的抗原性。 MAG_5040 的 UGA 校正版本缺乏编码信号肽的区域，在大肠杆菌中表达为 GST 融合蛋白。重组 GST-MAG_5040 表现出与典型的糖非特异性核酸内切酶和核酸外切酶类似的核酸酶活性，以 DNA 作为首选底物，在 20 mM MgCl2 存在下、温度范围为 37 至 45°C 时具有最佳活性。根据计算机分析，在大多数属于人型支原体组的已测序支原体中，编码 MAG_5040 的基因的位置始终位于 ABC 转运蛋白的上游。在无乳分枝杆菌中，MAG_5040 与 ABC 转运蛋白成分以及 MAG_5030 一起转录为多顺反子 RNA，通过 3D 建模和同源性搜索预测 MAG_5030 是糖溶质结合蛋白。在绵羊和山羊感染的自然模型中，感染后 9 个月内检测到抗 MAG_5040 抗体。考虑到其酶活性，MAG_5040可能在无乳支原体在宿主体内的存活中发挥关键作用，从而有助于宿主致病性。 MAG_5040 的鉴定为开发控制小反刍动物传染性无乳症的合适工具开辟了新的视角。
In this study the enzymatic activity of Mycoplasma agalactiae MAG_5040, a magnesium-dependent nuclease homologue to the staphylococcal SNase was characterized and its antigenicity during natural infections was established. A UGA corrected version of MAG_5040, lacking the region encoding the signal peptide, was expressed in Escherichia coli as a GST fusion protein. Recombinant GST-MAG_5040 exhibits nuclease activity similar to typical sugar-nonspecific endo- and exonucleases, with DNA as the preferred substrate and optimal activity in the presence of 20 mM MgCl2 at temperatures ranging from 37 to 45°C. According to in silico analyses, the position of the gene encoding MAG_5040 is consistently located upstream an ABC transporter, in most sequenced mycoplasmas belonging to the Mycoplasma hominis group. In M. agalactiae, MAG_5040 is transcribed in a polycistronic RNA together with the ABC transporter components and with MAG_5030, which is predicted to be a sugar solute binding protein by 3D modeling and homology search. In a natural model of sheep and goats infection, anti-MAG_5040 antibodies were detected up to 9 months post infection. Taking into account its enzymatic activity, MAG_5040 could play a key role in Mycoplasma agalactiae survival into the host, contributing to host pathogenicity. The identification of MAG_5040 opens new perspectives for the development of suitable tools for the control of contagious agalactia in small ruminants.
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