Recombinant Expression in Pichia pastoris System of Three Potent Kv1.3 Channel Blockers: Vm24, Anuroctoxin, and Ts6.
Recombinant Expression in Pichia pastoris System of Three Potent Kv1.3 Channel Blockers: Vm24, Anuroctoxin, and Ts6.
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DOI：
10.3390/jof8111215
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发表时间：
2022-11-17
期刊：
影响因子：
--
通讯作者：
Panyi G
中科院分区：
文献类型：
--
作者：
Borrego J;Naseem MU;Sehgal ANA;Panda LR;Shakeel K;Gaspar A;Nagy C;Varga Z;Panyi G
Kv1.3通道已成为治疗各种疾病的治疗靶点。几种Kv1.3通道阻滞剂已从蝎子毒液中分离出来;然而，广泛的研究需要大量的毒素，而这些毒素不能直接从毒液中获得。巴斯德毕赤酵母表达系统为克服化学合成和大肠杆菌表达的局限性提供了一种经济有效的方法。coli重组表达。在这项工作中，我们开发了一种有效的系统，用于从不同的蝎子毒液中生产三种有效的Kv1.3通道阻断剂：Vm 24，AnTx和Ts 6。使用毕赤酵母系统，这些毒素可以以足够的量（Vm 24 1.6 mg/L，AnTx 46 mg/L，和Ts 6 7.5 mg/L）获得以表征其生物活性。在标记的和未标记的重组肽的活性之间进行比较。标记的Vm 24和未标记的AnTx在阻断Kv1.3方面几乎等同于天然毒素（分别为Kd = 4.4 pM和Kd = 0.72 nM），而与天然肽相比，未标记的Ts 6表现出Kd增加53倍（Kd = 29.1 nM）。本文描述的方法提供了一种可以优化毒素生产的方法，以开发具有治疗潜力的更具选择性和有效性的Kv1.3阻断剂。
The Kv1.3 channel has become a therapeutic target for the treatment of various diseases. Several Kv1.3 channel blockers have been characterized from scorpion venom; however, extensive studies require amounts of toxin that cannot be readily obtained directly from venoms. The Pichia pastoris expression system provides a cost-effective approach to overcoming the limitations of chemical synthesis and E. coli recombinant expression. In this work, we developed an efficient system for the production of three potent Kv1.3 channel blockers from different scorpion venoms: Vm24, AnTx, and Ts6. Using the Pichia system, these toxins could be obtained in sufficient quantities (Vm24 1.6 mg/L, AnTx 46 mg/L, and Ts6 7.5 mg/L) to characterize their biological activity. A comparison was made between the activity of tagged and untagged recombinant peptides. Tagged Vm24 and untagged AnTx are nearly equivalent to native toxins in blocking Kv1.3 (Kd = 4.4 pM and Kd = 0.72 nM, respectively), whereas untagged Ts6 exhibits a 53-fold increase in Kd (Kd = 29.1 nM) as compared to the native peptide. The approach described here provides a method that can be optimized for toxin production to develop more selective and effective Kv1.3 blockers with therapeutic potential.
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影响因子：
3.7
作者：
Anangi R;Koshy S;Huq R;Beeton C;Chuang WJ;King GF
通讯作者：
King GF
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3.6
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2.8
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Bruce, C.;Fitches, E. C.;Gatehouse, J. A.
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影响因子：
4.1
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Booth WT;Schlachter CR;Pote S;Ussin N;Mank NJ;Klapper V;Offermann LR;Tang C;Hurlburt BK;Chruszcz M
通讯作者：
Chruszcz M