School of Life Science and Bio-pharmaceutics, Shenyang Pharmaceutical University, Shenyang, Liaoning Province 110016, PR China.
Peptides. 2010 Jun;31(6):995-1000. doi: 10.1016/j.peptides.2010.03.017. Epub 2010 Mar 20.
In this study, an effective Escherichia coli expression system was used to study the role of residues in the antitumor-analgesic peptide from Chinese scorpion Buthus martensii Karsch (BmKAGAP). To evaluate the extent to which residues of the toxin core contribute to its analgesic activity, nine mutants of BmKAGAP were obtained by PCR. Using site-directed mutagenesis, all of these residues were individually substituted by one amino acid. These were then subjected to a circular dichroism analysis, and an analgesic activity assay in mice. This study represents a thorough mapping and elucidation of the epitopes that underlie the molecular basis of the analgesic activity. The three-dimensional structure of BmKAGAP was established by homology modeling. Our results revealed large mutant-dependent differences that indicated important roles for the studied residues. With our ongoing efforts for establishing the structure and analgesic activity relationship of BmKAGAP, we have succeeded in pinpointing which residues are important for the analgesic activity.
在这项研究中,我们使用了一种有效的大肠杆菌表达系统来研究中国蝎子(Buthus martensii Karsch)抗肿瘤-镇痛肽残基的作用。为了评估毒素核心残基对其镇痛活性的贡献程度,我们通过 PCR 获得了 BmKAGAP 的 9 个突变体。通过定点突变,我们将这些残基分别突变为一个氨基酸。然后,我们对这些突变体进行圆二色性分析和小鼠镇痛活性测定。这项研究全面分析并阐明了构成镇痛活性分子基础的表位。通过同源建模建立了 BmKAGAP 的三维结构。我们的结果显示,突变体依赖性差异很大,表明研究的残基起着重要作用。通过我们正在进行的关于建立 BmKAGAP 结构和镇痛活性关系的研究,我们成功地确定了哪些残基对镇痛活性很重要。