Structural Studies and Char. of Neurotoxic Venom Peptides from Sicariidae Spiders

描述（由申请人提供）：蜘蛛毒液是包含数百种成分的复杂混合物，包括神经毒性肽和蛋白质。蜘蛛毒液固有的复杂性和大量的蜘蛛种类（约42000种已被描述）的结合意味着蜘蛛毒液中含有大量的毒素多样性，而这些毒素的多样性才刚刚开始被探索。本研究的目的是从褐隐蛛及其近缘蜘蛛（隐蛛科蜘蛛）的毒液中发现有趣的肽和蛋白质，并对这些肽和蛋白质的结构和功能进行表征。我们已经通过对粗毒液的色谱分离以及对蛇毒腺mRNA合成的cDNA文库的分析，探索了刺蛛科蜘蛛毒液成分的多样性。这有助于我们开始了解这些毒素的分子进化模式，并确定肽以供进一步研究。在Specific Aim 1中，将使用重组DNA技术在细菌中表达感兴趣的肽，以产生样品以进一步进行结构和功能表征。结构表征将使用NMR（核磁共振）光谱进行。在专项目标2中，这些肽的毒性和作用方法将被表征。本研究旨在描述这些毒液成分的结构和功能，以发现新颖性和结构进化分析为目标。蜘蛛毒液多肽和蛋白质的潜在用途是作为治疗药物和神经生理学研究的工具，因为它们中的许多被认为可以特异性地抑制或激活神经细胞中的离子通道和受体。此外，一些毒液肽具有物种特异性，因此有可能被用作环境友好型杀虫毒素。

DESCRIPTION (provided by applicant): Spider venoms are complex mixtures that contain hundreds of components, including neurotoxic peptides and proteins. The combination of the inherent complexity of spider venoms and the large number of spider species (~42,000 described) means that spider venoms contain a large array of toxin diversity that has only just begun to be explored. The aim of this research is to discover interesting peptides and proteins from the venom of the brown recluse spider and its relatives (the Sicariidae spiders), and then to characterize the structure and function of these peptides and proteins. We have previously explored the diversity of venom components from Sicariidae spiders by chromatographic separation of crude venom as well as by the analysis of cDNA libraries created from venom gland mRNA. These have helped us begin to understand the patterns of molecular evolution of these toxins and identify peptides for further study. In Specific Aim 1, interesting peptides will e expressed in bacteria using recombinant DNA techniques to generate samples for further structural and functional characterization. Structural characterization will be carried out using NMR (nuclear magnetic resonance) spectroscopy. In Specific Aim 2, the toxicity and method of action of these peptides will be characterized. The proposed research aims to characterize the structure and function of these venom components with the goals of discovery of novelty and analyses of structural evolution. Spider venom peptides and proteins are of interest for their potential use as therapeutic drugs and as tools for neurophysiology research, as many of them are believed to specifically inhibit or activate ion channels and receptors in nerve cells. In addition, some venom peptides are species-specific and therefore have the potential to be used as environmentally-friendly insecticidal toxins. PUBLIC HEALTH RELEVANCE: The proposed research aims to determine the structure and function of neurotoxic peptides and proteins identified from spider venoms. These components of spider venom are of interest for their potential use as therapeutic drugs and as tools for neurophysiology research, as many of them are believed to specifically inhibit or activate ion channels and receptors in nerve cells. In addition, some venom peptides are species-specific and therefore have the potential to be used as environmentally-friendly insecticidal toxins.