| 鸡毒支原体磷酸酶PrpC活性检测 |
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| 引用本文： | 胡美容,陈鸿军,于圣青,仇旭升,谭磊,高崧,丁铲.鸡毒支原体磷酸酶PrpC活性检测[J].动物医学进展,2012(9):1-5. |
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| 作者姓名： | 胡美容 陈鸿军 于圣青 仇旭升 谭磊 高崧 丁铲 |
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| 作者单位： | 1. 中国农业科学院上海兽医研究所,上海200241
2. 扬州大学兽医学院,江苏扬州225009 |
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| 基金项目： | 国家自然科学基金项目(30871883,31001077) |
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| 摘 要： | HPr激酶/磷酸酶(PrkC/PrpC)系统在细菌和支原体中高度保守,不仅参与糖酵解酶类的磷酸化/去磷酸化,也与毒力、生物被膜等生命活动相关。克隆并突变获得编码鸡毒支原体磷酸酶PrpC的ptc1基因,经原核表达纯化后,利用底物PNPP体外检测其酶活性,并对影响该酶活性的pH、金属离子、温度等影响因子进行分析。结果表明,Ptc1蛋白具有磷酸酶活性,Mn2+为该酶辅因子,最适离子浓度为2mol/L,最适pH为8.5,最适温度为37℃。相同浓度的Li+、Na+、K+、Mg2+、Ca2+、Ba2+、Al 3+、Hg2+等金属离子均对表达产物具有不同程度的抑制活性。初步建立了PrpC功能检测体系,为进一步深入研究PrkC/PrpC调节系统奠定基础。
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| 关 键 词： | 鸡毒支原体 PrpC 磷酸酶 |
Activity Detection Characteristic of Phosphatase PrpC in Mycoplasma gallisepticum |
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| HU Mei-rong,CHEN Hong-jun,YU Sheng-qing,QIU Xu-sheng,TAN Lei,GAO Song,DING Chan.Activity Detection Characteristic of Phosphatase PrpC in Mycoplasma gallisepticum[J].Progress In Veterinary Medicine,2012(9):1-5. |
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| Authors: | HU Mei-rong CHEN Hong-jun YU Sheng-qing QIU Xu-sheng TAN Lei GAO Song DING Chan |
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| Institution: | 1(1.Shanghai Veterinary Research Institute,Chinese Academy of Agricultural Science,Shanghai,200241,China; 2.Veterinary Medicine College of Yangzhou University,Yangzhou,Jiangsu,225009,China) |
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| Abstract: | HPr kinase/phosphatase(PrkC/PrpC) system is highly conserved in bacteria and mycoplasma,it involves not only in the glycolytic enzymes of phosphorylation/dephosphorylation,but in life activities,such as virulence and biofilm.In this study,the ptc1 gene of Mycoplasma gallisepticum Rlow was cloned and expressed in Escherichia coli BL21(DE3).Expressed PrpC protein was purified and used to catalyze PNPP dephosphorylation.The optimal pH,concentration of metal ions and temperature were investigated in vitro.The results showed that PrpC exhibited its maximal activity in pH8.5 reaction buffer containing 2 mmol/L Mn2+ at 37℃.The PrpC enzyme is a kind of Mn2+-dependent dephosphatase.However,1 mmol/L of Li+,Na+,K+,Mg2+,Ca2+,Ba2+,Al3+ and Hg2+ could inhibited the enzymatic activity.The study established initially PrpC function testing system which would provide a powerful means in studying the PrkC/PrpC regulation systems in the further. |
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| Keywords: | Mycoplasma gallisepticum PrpC phosphatase |
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