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

描述（由申请人提供）：蜘蛛毒液含有数百种成分，包括神经毒性肽和蛋白质。这些毒液成分因其作为治疗药物和神经生理学研究工具的潜在用途而引起人们的兴趣，因为它们中的许多成分特异性地抑制或激活神经细胞中的离子通道和受体。此外，一些毒液肽是物种特异性的。例如，某些肽只影响昆虫，但对哺乳动物无害；因此，蜘蛛毒液肽有可能作为一种环保型杀虫毒素加以利用。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。在本研究中，我们将通过对粗毒液的色谱分离以及对由毒液腺mRNA构建的cDNA文库的分析，来探讨刺蛛科蜘蛛毒液成分的多样性。这将有助于了解这些毒素的分子进化模式，并为进一步研究鉴定肽。在Specific Aim 2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。最后，在特异性目标3中，这些肽的毒性和作用方法将被表征。通过鉴定和表达毒液肽和蛋白质，本研究旨在表征这些毒液成分的结构和功能，以发现新颖性和分析结构进化。公共卫生相关性：拟议的研究旨在从蜘蛛毒液中识别神经毒性肽和蛋白质，然后确定其结构和功能。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的杀虫毒素。

DESCRIPTION (provided by applicant): Spider venoms contain hundreds of components, including neurotoxic peptides and proteins. These venom components are of interest for their potential use as therapeutic drugs and as tools for neurophysiology research, as many of them specifically inhibit or activate ion channels and receptors in nerve cells. In addition, some venom peptides are species-specific. For example, certain peptides only affect insects but are harmless to mammals; consequently, it is possible that spider venom peptides could be used as environmentally-friendly insecticidal toxins. 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. In Specific Aim 1, the diversity of venom components from Sicariidae spiders will be explored by chromatographic separation of crude venom as well as by the analysis of cDNA libraries created from venom gland mRNA. This will help in understanding the patterns of molecular evolution of these toxins and identify peptides for further study. In Specific Aim 2, interesting peptides will be 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. Finally, in Specific Aim 3, the toxicity and method of action of these peptides will be characterized. By identifying and then expressing venom peptides and proteins 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. PUBLIC HEALTH RELEVANCE: The proposed research aims to identify neurotoxic peptides and proteins from spider venoms and then to determine their structure and function. 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.