A serine/threonine phosphatase encoded by MG_207 of Mycoplasma genitalium is critical for its virulence.
A serine/threonine phosphatase encoded by MG_207 of Mycoplasma genitalium is critical for its virulence.
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DOI：
10.1186/1471-2180-13-44
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发表时间：
2013-02-21
期刊：
影响因子：
4.2
通讯作者：
Dhandayuthapani S
中科院分区：
文献类型：
--
作者：
Martinez MA;Das K;Saikolappan S;Materon LA;Dhandayuthapani S
细菌信号转导系统如双组分系统（TCS）、丝氨酸/苏氨酸激酶（STK）和丝氨酸/苏氨酸磷酸酶（STP）在细菌致病和致病过程中起着重要作用。生殖支原体是一种分别引起男性和女性泌尿生殖系统疾病尿道炎和宫颈炎的柔膜菌，是一种缺乏TCS但具有STK/STP的病原体。在这项研究中，我们调查了该物种的基因MG_207编码的STP蛋白的生化和毒力特性。我们在大肠杆菌过表达系统中过表达MG 207作为重组His 10 MG 207蛋白，并通过亲和层析纯化。该重组蛋白以剂量依赖性方式容易地水解底物对硝基苯基磷酸盐（pNPP）。使用合成肽作为底物的其他研究表明，重组蛋白能够水解磷酸苏氨酸。此外，M.当与野生型G37菌株相比时，缺乏蛋白MG 207的生殖器（TIM 207）显示差异磷酸化的蛋白。质谱分析结果表明，TIM 207菌株中磷酸化差异的关键蛋白可能是MG_328基因编码的细胞骨架蛋白和MG_274基因编码的丙酮酸脱氢酶E1 α链。此外，注意到TIM 207对HeLa细胞的细胞毒性较小，这与该菌株产生较少的过氧化氢有关。与野生型M相比，该菌株在诱导THP-1细胞系分化方面的效率也较低。生殖器研究结果提示MG 207是M.生殖器及其存在可能对该物种的毒力至关重要。
Bacterial signal transduction systems like two component system (TCS) and Serine/Threonine kinase (STK) and Serine/Threonine phosphatase (STP) play important roles in the virulence and pathogenesis of bacterial pathogens. Mycoplasma genitalium, a mollicute that causes the urogenital diseases urethritis and cervicitis in men and women, respectively, is a pathogen which lacks TCS but possesses STK/STP. In this study, we investigated the biochemical and virulence properties of an STP protein encoded by the gene MG_207 of this species. We overexpressed MG207 in Escherichia coli overexpression system as a recombinant His10MG207 protein and purified it with affinity chromatography. This recombinant protein readily hydrolyzed the substrate p-nitrophenyl phosphate (pNPP) in a dose-dependent manner. Additional studies using synthetic peptides as substrates revealed that the recombinant protein was able to hydrolyze the threonine phosphate. Further, a transposon insertion mutant strain of M. genitalium (TIM207) that lacks the protein MG207 showed differentially phosphorylated proteins when compared to the wild type G37 strain. Mass spectrometry revealed that some of the key proteins differentially phosphorylated in TIM207 strain were putative cytoskeletal protein encoded by the gene MG_328 and pyruvate dehydrogenase E1 α chain encoded by the gene MG_274. In addition, TIM207 was noticed to be less cytotoxic to HeLa cells and this correlated with the production of less hydrogen peroxide by this strain. This strain was also less efficient in inducing the differentiation of THP-1 cell line as compared to wild type M. genitalium. The results of the study suggest that MG207 is an important signaling protein of M. genitalium and its presence may be crucial for the virulence of this species.
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影响因子：
3.2
作者：
Dhandayuthapani, S;Blaylock, MW;Baseman, JB
通讯作者：
Baseman, JB
影响因子：
6.8
作者：
Barford, D
通讯作者：
Barford, D
影响因子：
3.2
作者：
Dhandayuthapani, S;Mudd, M;Deretic, V
通讯作者：
Deretic, V
影响因子：
56.9
作者：
FRASER, CM;GOCAYNE, JD;VENTER, JC
通讯作者：
VENTER, JC
影响因子：
3.1
作者：
Echenique, J;Kadioglu, A;Trombe, MC
通讯作者：
Trombe, MC