Molecular cloning, bioinformatics analysis and functional characterization of HWTX-XI toxin superfamily from the spider Ornithoctonus huwena
Molecular cloning, bioinformatics analysis and functional characterization of HWTX-XI toxin superfamily from the spider Ornithoctonus huwena
复制标题
虎纹鸟蛛 HWTX-XI 毒素超家族的分子克隆、生物信息学分析及功能表征
DOI：
10.1016/j.peptides.2014.01.001
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发表时间：
2014-04-01
期刊：
影响因子：
3
通讯作者：
Liang, Songping
中科院分区：
文献类型：
--
作者：
Jiang, Liping;Deng, Meichun;Liang, Songping
蜘蛛毒液含有非常有价值的天然资源库，以发现用于分子多样性分析和治疗应用的新组分。本研究对虎纹捕鸟蛛（Ornithoctonus huwena）蜘蛛毒腺中的Kunitz型毒素HWTX-XI进行了直接克隆、分析和功能鉴定。迄今为止，HWTX-XI超家族由121个高质量的表达序列标签推导出的38个成员组成，是最大的蜘蛛KU超家族，其分子多样性主要来自cDNA串联重复序列和局部超突变。其中，HW 11 c40和HW 11 c50可能是天然Kunitz毒素和sub-Kunitz毒素之间的中间变体。为了阐明其生物学活性，成功表达了重组蛋白HW 11 c4、HW 11 c24、HW 11 c27和HW 11 c39，并对其进行了纯化和功能鉴定。HW 11 c4和HW 11 c27都显示出对胰蛋白酶、胰凝乳蛋白酶和激肽释放酶的抑制活性。此外，HW 11 c4也是Kv1.1通道的相对特异性抑制剂。发现HW 11 c24和HW 11 c39对糜蛋白酶、胰蛋白酶、激肽释放酶、凝血酶和离子通道无活性。这些发现为HWTX-XI超家族的毒素多样化提供了分子证据，并为开发潜在的临床应用提供了有用的丝氨酸蛋白酶抑制剂和离子通道阻断剂的分子模板。(c)2014爱思唯尔公司All rights reserved.
Spider venom contains a very valuable repertoire of natural resources to discover novel components for molecular diversity analyses and therapeutic applications. In this study, HWTX-XI toxins from the spider venom glands of Ornithoctonus huwena which are Kunitz-type toxins (KTTs) and were directly cloned, analyzed and functionally characterized. To date, the HWTX-XI superfamily consists of 38 members deduced from 121 high-quality expressed sequence tags, which is the largest spider KU superfamily with significant molecular diversity mainly resulted from cDNA tandem repeats as well as focal hypermutation. Among them, HW11c40 and HW11c50 may be intermediate variants between native Kunitz toxins and sub-Kunitz toxins based on evolutionary analyses. In order to elucidate their biological activities, recombinant HW11c4, HW11c24, HW11c27 and HW11c39 were successfully expressed, further purified and functionally characterized. Both HW11c4 and HW11c27 display inhibitory activities against trypsin, chymotrypsin and kallikrein. Moreover, HW11c4 is also an inhibitor relatively specific for Kv1.1 channels. HW11c24 and HW11c39 are found to be inactive on chymotrysin, trypsin, kallikrein, thrombin and ion channels. These findings provide molecular evidence for toxin diversification of the HWTX-XI superfamily and useful molecular templates of serine protease inhibitors and ion channel blockers for the development of potentially clinical applications. (c) 2014 Elsevier Inc. All rights reserved.