FULL CHARACTERIZATION OF CONUS VENOM BY CHROMATOGRAPHY AND MASS SPECTROMETRY

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 芋螺毒素是芋螺蛇毒中的生物活性成分。圆锥蜗牛是一大类有毒腹足类动物，约有500个物种。所有锥形蜗牛物种都是通过注射毒液来捕获或杀死猎物的捕食者。该物种被细分为食鱼(食鱼)、食软体动物(食软体动物)和食虫(食蠕虫)。有趣的是，圆锥蛇毒具有显著多样性的生物活性神经肽。他们的目标是神经肌肉系统中的离子通道和受体。例如，地理圆锥蛇毒含有作用于电压敏感钙通道、钠通道和N-甲基-D-天冬氨酸(NMDA)受体的高亲和力多肽(螺毒素)。虽然芋螺毒素的研究已经有30多年的历史，但在蛋白质水平上还只鉴定出某些物种的少数毒素成分。这个项目的目的之一是绘制出来自文氏圆锥和圆锥纺织的毒液的巨大复杂性。我们将利用质谱学进行蛋白质和多肽的鉴定。具有生物功能的多肽通常含有二硫键(半胱氨酸)，通过两个半胱氨酸残基的游离硫醇基团连接起来。二硫键的形成是最常见的翻译后修饰之一。二硫化物对维持或建立蛋白质和多肽的三维结构很重要，也参与调节多肽的活性。高二硫键桥联多肽的一个例子是芋螺毒素。典型的这些毒素由20到30个氨基酸组成，含有2到3个二硫化物。该项目的第二个主要目标是指定芋螺毒素中的二硫化物。利用层析和质谱法可以首先从圆锥蛇毒中提纯和分离芋螺毒素，并利用串联质谱法绘制半胱氨酸支架图和确定半胱氨酸连接性。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Conotoxins are the bio-active components of Conus snail venom. Conus snails are a large genus of venomous gastropods comprising approximately 500 species. All conus snail species are predators that inject venom to capture or kill prey. The species are subdivided in fish eaters (piscivorous), mollusc eaters (molluscivorous) and worm eaters (vermivorous). Interestingly, Conus venoms have a remarkable diversity of biological active neuropeptides. Their targets are ion channels and receptors in the neuromuscular system. For instance, the venom of Conus Geographus contains high affinity peptides (conotoxins) that act on voltage sensitive calcium channels, sodium channels and N-methyl-D-aspartate (NMDA) receptors. Although conotoxins have been studied for over 30 years, only a few components of the venoms of some species have been identified on the protein level. One of the aims of this project is to map the enormous complexity of the venom derived from both Conus Ventricosus and Conus Textile. We will utilize mass spectrometry for protein and peptide assignment. Peptides with biological functions often contain disulfide bridges (cystines) connecting two cysteine residues via their free thiol groups. Disulfide bond formation is one of the most common posttranslational modification. Disulfides are important to maintain or establish the 3D structure of proteins and peptides and are also involved in regulating peptide activity. An example of highly disulfide bridged peptides are conotoxins. Typically these toxins are 20 to 30 amino acids long containing 2 or 3 disulfides. A secondairy aim of this project is the assignment of disulfides in conotoxins. Chromatography and mass spectrometry can be used to first purify and isolate conotoxins from Conus venom and map the cysteine scaffold and determine the cystine connectivity utilizing tandem mass spectrometry.