Identification and Characterization of the Nuclease Activity of the Extracellular Proteins from Salmonella enterica Serovar Typhimurium
Identification and Characterization of the Nuclease Activity of the Extracellular Proteins from Salmonella enterica Serovar Typhimurium
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鼠伤寒沙门氏菌胞外蛋白核酸酶活性的鉴定和表征
DOI：
10.1007/s00284-020-02201-1
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发表时间：
2020-09
影响因子：
2.6
通讯作者：
Wu Tingcai
中科院分区：
文献类型：
--
作者：
Liao Chengshui;Zhang Mengke;Cheng Xiangchao;Li Qi;Mao Fuchao;Wang Xiaoli;Yu Chuan;Yu Zuhua;Jia Yanyan;Li Jing;He Lei;Zhang Chunjie;Li Yinju;Wu Tingcai
病原体已经进化出一系列策略来建立生产性感染。病原体分泌的胞外蛋白是其逃避宿主天然免疫应答的独特机制之一。沙门氏菌分泌系统转运的许多分泌蛋白已被广泛研究。某些细胞外核酸酶对于细菌的发病机理是必不可少的。然而，目前还没有关于沙门氏菌分泌的蛋白质的酶性质的数据。因此，在本研究中，我们已经确定和表征的核酸酶活性的胞外蛋白从沙门氏菌肠血清型鼠伤寒沙门氏菌。结果表明，S.琼脂糖凝胶电泳和琼脂平板扩散法显示鼠伤寒沙门氏菌对λDNA具有脱氧核糖核酸酶活性。在16 °C、37 °C和42 °C下观察到活性，发现在42 °C下最高，在超过60 °C的温度下受到抑制。该酶在碱性条件下（pH 7-10）活性稳定，最适pH为9.0。高离子强度的Ba ~（2+）、Ca ~（2+）、Mg ~（2+）和Ni ~（2+）对核酸酶活性有促进作用。核酸酶谱分析显示，胞外蛋白有4条活性带; LC-ESI/MS/MS分析鉴定出7条蛋白。核酸酶谱分析表明，原核表达的重组5′-核苷酸酶蛋白具有DNase活性。据我们所知，目前的研究结果代表了第一个直接和明确的证明核酸酶活性的胞外蛋白从S。鼠伤寒沙门氏菌，为进一步研究细胞外蛋白在致病性和免疫逃避中的作用提供了重要的基础。
Pathogens have evolved an array of strategies to establish a productive infection. The extracellular proteins secreted by pathogens are one of unique mechanisms to evade the host innate immune response. Many secretory proteins transported by the bacterial secretion systems have been widely investigated in Salmonella. Certain extracellular nucleases are essential for bacterial pathogenesis. However, there is no current data available for the enzymatic properties of the proteins secreted by Salmonella. Therefore, in the present study we have identified and characterized the nuclease activity of the extracellular proteins from Salmonella enterica serovar Typhimurium. It was demonstrated that the extracellular proteins from S. Typhimurium exhibited the deoxyribonucleases activity against λDNA by agarose gel electrophoresis and agar plate diffusion method. The activity was observed at 16 °C, 37 °C and 42 °C, and found to be highest at 42 °C and inhibited at temperatures over 60 °C. The nuclease activity was stable under alkaline conditions (pH 7–10) and the optimum pH was 9.0. The nuclease activity was promoted at high ionic strength of Ba2+, Ca2+, Mg2+, and Ni2+. Nuclease zymography analysis revealed that there were four activity bands in the extracellular proteins; followed by LC-ESI/MS/MS analysis seven proteins were identified. As demonstrated by nuclease zymography, the recombinant 5′-nucleotidase protein expressed in the prokaryotic expression system displayed the DNase activity. To our knowledge, the present findings represent the first direct and unambiguous demonstration of the nuclease activity of the extracellular proteins from S. Typhimurium, and it provides an important fundamental for further investigation of the role of the extracellular proteins in pathogenicity and immune evasion.
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来自变形链球菌的核酸酶促进生物膜分散并逃避中性粒细胞胞外陷阱的杀伤
影响因子：
5.7
作者：
Liu J;Sun L;Liu W;Guo L;Liu Z;Wei X;Ling J
通讯作者：
Ling J
影响因子：
3.7
作者：
Doke M;Fukamachi H;Morisaki H;Arimoto T;Kataoka H;Kuwata H
通讯作者：
Kuwata H
影响因子：
5.2
作者：
Malik-Kale P;Jolly CE;Lathrop S;Winfree S;Luterbach C;Steele-Mortimer O
通讯作者：
Steele-Mortimer O
DOI：
10.1093/jb/mvy039
发表时间：
2018-08
期刊：
The Journal of Biochemistry
影响因子：
--
作者：
Kar Yan Soh;J. Loh;T. Proft
通讯作者：
Kar Yan Soh;J. Loh;T. Proft
影响因子：
2.4
作者：
DETWILER, C;MACINTYRE, R
通讯作者：
MACINTYRE, R