Lin Shengguo, Wang Xuelin, Hu Xueyao, Zhao Yongshan, Zhao Mingyi, Zhang Jinghai, Cui Yong
School of Life Science and Bio-pharmaceutics, Shenyang Pharmaceutical University, Shenyang, Liaoning Province 110016, China.
School of Medical Devices, Shenyang Pharmaceutical University, Benxi, Liaoning Province 117004, China.
Protein Pept Lett. 2017;24(3):235-240. doi: 10.2174/0929866524666170117142404.
Scorpion venom contains a large variety of biologically active peptides. However, most of these peptides have not been identified and characterized. Peptides with three disulfide bridges, existing in the scorpion venom, have not been studied in detail and have been poorly characterized until now. Here, we report the recombinant expression and functional characterization of two kinds of venom peptides (BmKBTx and BmNaL-3SS2) with three disulfide bridges. This study adopted an effective Escherichia coli system. The genes for BmKBTx and BmNaL-3SS2 were obtained by polymerase chain reaction and cloned to the pSYPU-1b vector. After expression and purification, the two recombinant proteins were subjected to an analgesic activity assay in mice and whole-cell patchclamp recording of hNav1.7-CHO cell lines. Functional tests showed that BmKBTx and BmNaL- 3SS2 have analgesic activity in mice and can interact with the hNav1.7 subtype of the voltage-gated sodium channel (VGSC). Scorpion venom is rich in bioactive proteins, but most of their functions are unknown to us. This study has increased our knowledge of these novel disulfide-bridged peptides (DBPs) and their biological activities.
蝎毒含有多种生物活性肽。然而，这些肽中的大多数尚未被鉴定和表征。存在于蝎毒中的具有三个二硫键的肽，到目前为止尚未得到详细研究，其特征也了解甚少。在此，我们报告了两种具有三个二硫键的毒液肽（BmKBTx和BmNaL-3SS2）的重组表达及功能表征。本研究采用了一种有效的大肠杆菌系统。通过聚合酶链反应获得了BmKBTx和BmNaL-3SS2的基因，并将其克隆到pSYPU-1b载体中。表达和纯化后，对这两种重组蛋白进行了小鼠镇痛活性测定以及hNav1.7-CHO细胞系的全细胞膜片钳记录。功能测试表明，BmKBTx和BmNaL-3SS2在小鼠中具有镇痛活性，并且可以与电压门控钠通道（VGSC）的hNav1.7亚型相互作用。蝎毒富含生物活性蛋白，但其大多数功能我们尚不清楚。这项研究增加了我们对这些新型二硫键连接肽（DBP）及其生物活性的了解。
