Mycoplasma synoviae enolase is a plasminogen/fibronectin binding protein.
Mycoplasma synoviae enolase is a plasminogen/fibronectin binding protein.
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DOI：
10.1186/s12917-014-0223-6
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发表时间：
2014-09-25
影响因子：
2.6
通讯作者：
Ding C
中科院分区：
文献类型：
--
作者：
Bao S;Guo X;Yu S;Ding J;Tan L;Zhang F;Sun Y;Qiu X;Chen G;Ding C
滑膜支原体是一种禽类病原体，可引起鸡和火鸡的呼吸道感染和关节炎，给养禽业造成严重的经济损失。据报道，烯醇化酶在几种细菌病原体中起重要作用，但它在滑膜分枝杆菌中的作用尚未确定。因此，本研究从WVU1853菌株中扩增出滑膜分枝杆菌烯醇化酶编码基因(ENO)，并在大肠杆菌BL21细胞中表达。并对其酶活性、免疫原性及与鸡纤溶酶原(PLG)和人纤维连接蛋白(FN)的结合活性进行了评价。结果表明，重组滑膜菌烯醇化酶蛋白(RMsENO)能催化2-磷酸甘油酸酯(2-PGA)转化为磷酸烯醇式丙酮酸(PEP)，其Km值为1.1Mol× 10 3M，Vmax为0.739−/L/μ。免疫印迹和免疫电子显微镜分析证实，烯醇化酶分布在滑膜细胞的表面和胞浆内。结合实验表明，rMsEno能与鸡PLG和人FN蛋白结合。补体依赖的支原体杀灭试验表明，兔抗rMsENO血清在补体存在下具有明显的杀支原体效果，这也证实了烯醇化酶分布在滑膜表面。抑制实验表明，兔抗rMsENO血清能有效抑制经PLG处理的DF-1细胞与滑膜支原体的黏附。这些结果表明，滑膜分枝杆菌烯醇化酶对2-PGA转化为PEP具有良好的催化活性，并与鸡PLG和人FN结合。兔抗rMsENO血清具有明显的补体依赖的支原体杀灭作用和黏附抑制作用。这些结果表明滑膜分枝杆菌烯醇化酶在滑膜分枝杆菌的代谢中起着重要作用，并可能影响滑膜分枝杆菌的感染和免疫。本文的在线版本(doi：10.1186/s12917-0140223-6)包含补充材料，授权用户可以使用。
Mycoplasma synoviae is an avian pathogen that can lead to respiratory tract infections and arthritis in chickens and turkeys, resulting in serious economic losses to the poultry industry. Enolase reportedly plays important roles in several bacterial pathogens, but its role in M. synoviae has not been established. Therefore, in this study, the enolase encoding gene (eno) of M. synoviae was amplified from strain WVU1853 and expressed in E. coli BL21 cells. Then the enzymatic activity, immunogenicity and binding activity with chicken plasminogen (Plg) and human fibronectin (Fn) was evaluated. We demonstrated that the recombinant M. synoviae enolase protein (rMsEno) can catalyze the conversion of 2-phosphoglycerate (2-PGA) to phosphoenolpyruvate (PEP), the Km and Vmax values of rMsEno were 1.1 × 10−3 M and 0.739 μmol/L/min, respectively. Western blot and immuno-electron microscopy analyses confirmed that enolase was distributed on the surface and within the cytoplasm of M. synoviae cells. The binding assays demonstrated that rMsEno was able to bind to chicken Plg and human Fn proteins. A complement-dependent mycoplasmacidal assay demonstrated that rabbit anti–rMsEno serum had distinct mycoplasmacidal efficacy in the presence of complement, which also confirmed that enolase was distributed on the surface of M. synoviae. An inhibition assay showed that the adherence of M. synoviae to DF-1 cells pre-treated with Plg could be effectively inhibited by treatment with rabbit anti-rMsEno serum. These results reveal that M. synoviae enolase has good catalytic activity for conversion of 2-PGA to PEP, and binding activity with chicken Plg and human Fn. Rabbit anti–rMsEno serum displayed an obvious complement-dependent mycoplasmacidal effect and adherent inhibition effect. These results suggested that the M. synoviae enolase plays an important role in M. synoviae metabolism, and could potentially impact M. synoviae infection and immunity. The online version of this article (doi:10.1186/s12917-014-0223-6) contains supplementary material, which is available to authorized users.
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影响因子：
3.3
作者：
Menegatti, Angela C. O.;Tavares, Carolina P.;Terenzi, Hernan
通讯作者：
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