BmKANEPII同源模建及其活性位点预测.docVIP

Molecular Modeling and Active Sites Prediction of BmK ANEPII MA Lin, WANG Jian, YANG Wen-yao, ZHANG Jing-hai,SONG Yong-bo （School of Biological Science and Biopharmacy, Shenyang Pharmaceutical University，Shenyang, 110016）BmK ANEPII, a depressant insect toxin from Buthus martensii Karsch, has effect on sodium channels. In this paper, three-dimensional structure modeling of BmK ANEPII was carried out by the homology modeling with the software Modeller9v1, the initial model was refined by molecular mechanics and dynamics simulation by using Sybyl6.91 Software Package on the Silicon Graphics Inc.（SGI） Fuel work station．The final refined model is further assessed by Procheck to examine the quality of the model structures by checking the stability．Sequences and structures alignments between BmK ANEPII and some long-chain scorpion neurotoxins with analgesic effect were performed to indicate that they had the same conservative structures related to activity, which included multiple disulfide bonds，conserved aromatic residues and site C．The results obtained will be helpful for further experimental investigations． Key words：BmK ANEPII，homology modeling，analgesic effect BmK ANEPII同源模建及活性位点预测摘要：东亚钳蝎（Buthus martensii Karsch, BmK）抗神经兴奋活性肽（anti-neuroexcitation peptide II precursor, ANEPII）是作用于钠离子通道的一种重要长链蝎毒 。本文同源模建的方法运用Modeller9v1软件BmK ANEPII三维结构，Sybyl6.91软件包对初始模型进行分子力学优化用Procheck对模建结构进行合理性验证Site C区BmK ANEPII结构与镇痛活性之间的关系提供理论支持。 关键词：BmK ANEPII 同源模建 镇痛作用 yongbo7200@ Tel：024东亚钳蝎（Buthus martensii Karsch, BmK）抗神经兴奋活性肽（anti-neuroexcitation peptide II precursor, ANEPII）昆虫抑制性毒素。对该肽的研究包括分离纯化、分子克隆[1]、原核及真核[2]体系表达等，但对其结构与功能关系的研究还未见报道。BmK ANEPII除具有抗癫痫作用外，还具有良好的镇痛活性。这些研究结果引发了我们进一步研究BmK ANEPII结构与镇痛功能关系的兴趣。 最初人类对蛋白质的研究方法是X晶体衍射法。此方法的应用前提是能获得对X射线产生强衍射的晶体，其结构测定过程受晶体的表达、提纯等操作过程影响，晶体不稳定。而多维核磁共振技术则避免了这一困难。该方法能测定蛋白质的溶液结构，但目前为止仅适用于大约15种小蛋白左右，且消耗大，成本高。目前，人们多定点突变方法，盲目性。BmK ANEPII的结构进行模建及优化，并初步预测该肽的活性位点及活性区，为进一步实验研究BmK ANEPII结构与镇痛活性之间的关系提供理论支持。 材料与方法 材料 东亚钳蝎活性肽（BmK ANEPII）由85个氨基酸组成，其中前21个氨基酸是