Zhang Shaohua, You Yanan, Luo Xuenong, Zheng Yadong, Cai Xuepeng
State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, 730046, China.
State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, 730046, China.
Vet Parasitol. 2018 Apr 30;254:36-42. doi: 10.1016/j.vetpar.2018.02.041. Epub 2018 Feb 25.
Enolase (EC 4.2.1.11) acts as a multifunctional enzyme in many organisms, being involved in metabolism, transcription regulation and pathogenesis. In the current study, the recombinant α-enolase from Taenia solium (His-Tseno) was prepared and antiserum against His-Tseno was generated in rabbits. Consequently, we analyzed the enzymatic characteristics, plasminogen binding activity, tissue localization and expression patterns of Tseno. The study demonstrated that the enzymatic activity of His-Tseno was enhanced at pH around 7.0-7.5 and affected by addition of metal ions. Kinetic measurements using 2-phospho-d-glycerate (2-PGA) substrates gave a specific activity of 60.72 ± 0.84 U/mg and 1.1 mM of Km value. Plasminogen binding assay showed that His-Tseno could bind to human plasminogen and generate plasmin activated by a tissue-type plasminogen activator (t-PA). In addition, the lysine analogue 6-aminocaproic acid (ε-ACA) could inhibit the binding of plasminogen to His-Tseno. Quantitative real-time PCR confirmed that Tseno was expressed 2.38 folds higher in the adult worms (p < 0.05) than in the cysticerci. Further, an immunolocalization assay indicated that native Tseno was mainly distributed in the tegument and eggs of gravid proglottis from adult T. solium. In conclusion, Tseno executes the innate glycolytic function to supply energy for the growth, egg production, and even invasion of T. solium.
烯醇化酶（EC 4.2.1.11）在许多生物体中作为一种多功能酶，参与新陈代谢、转录调控和发病机制。在本研究中，制备了来自猪带绦虫的重组α-烯醇化酶（His-Tseno），并在兔体内产生了针对His-Tseno的抗血清。因此，我们分析了Tseno的酶学特性、纤溶酶原结合活性、组织定位和表达模式。研究表明，His-Tseno的酶活性在pH约7.0 - 7.5时增强，并受金属离子添加的影响。使用2-磷酸-d-甘油酸（2-PGA）底物进行的动力学测量得出比活性为60.72±0.84 U/mg，Km值为1.1 mM。纤溶酶原结合试验表明，His-Tseno可与人纤溶酶原结合，并产生由组织型纤溶酶原激活剂（t-PA）激活的纤溶酶。此外，赖氨酸类似物6-氨基己酸（ε-ACA）可抑制纤溶酶原与His-Tseno的结合。定量实时PCR证实，Tseno在成虫中的表达比囊尾蚴高2.38倍（p<0.05）。此外，免疫定位试验表明，天然Tseno主要分布在成年猪带绦虫妊娠节片的皮层和虫卵中。总之，Tseno执行先天性糖酵解功能，为猪带绦虫的生长、产卵甚至侵袭提供能量。
