Department of Biomolecular Sciences, Weizmann Institute of Science, Rehovot, Israel.
Department of Biomolecular Sciences, Weizmann Institute of Science, Rehovot, Israel.
Methods Enzymol. 2021;654:169-201. doi: 10.1016/bs.mie.2021.01.029. Epub 2021 Mar 9.
Animal venom is a rich source for peptide toxins that bind and modulate the function of ion channels. Owing to their ability to bind receptor sites on the channel protein with high affinity and specificity, peptide neurotoxins have become an indispensable tool for ion channel research. Recent breakthroughs in structural biology and advances in computer simulations of biomolecules have sparked a new interest in animal toxins as probes of channel protein structure and function. Here, we focus on methods used to produce animal toxins for research purposes using recombinant expression. The specific challenges associated with heterologous production of venom peptides are discussed, and several methods targeting these issues are presented with an emphasis on E. coli based systems. An efficient protocol for the bacterial expression, folding, and purification of recombinant venom peptides is described.
动物毒液是富含肽毒素的资源,这些毒素可以结合并调节离子通道的功能。由于其能够以高亲和力和特异性结合通道蛋白上的受体位点,肽神经毒素已成为离子通道研究不可或缺的工具。结构生物学的最新突破和生物分子计算机模拟的进展,激发了人们对动物毒素作为通道蛋白结构和功能研究探针的新兴趣。在这里,我们重点介绍使用重组表达生产用于研究目的的动物毒素的方法。讨论了异源生产毒液肽所面临的具体挑战,并提出了几种针对这些问题的方法,重点是基于大肠杆菌的系统。本文描述了一种用于细菌表达、折叠和纯化重组毒液肽的有效方案。