CHARACTERIZING TEREBRIDAE TOXINS

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 该项目的目的是识别Terebridae和角塔物种的毒素。在Orbitrap上用LC-MS法测定了石斑鱼(Terebridae Gutatta)、银鱼(Agyosia)、毛虾(Maunti)和福尔摩沙(Formosa)粗毒中存在的多肽。只有杜仲胶和银鱼毒液有足够的物质可以离线分离。 用反相高效液相色谱分离这两种毒素，并用MALDI TOF质谱仪对在214 nm处有较强紫外吸收的部分进行分析，以确定这些部分中潜在毒素的二硫键的准确质量和数目。 紫外可见光谱分析表明，绿背天牛毒液几乎没有显示任何物质，而杜仲胶天牛毒液有5个主要峰。不幸的是，使用标准的还原方案，所有的瓜塔多肽都会沉淀下来。通过增加有机溶剂(乙腈和甲醇)的用量和添加GnHCl对还原方案进行了优化。鉴定出质量在3700~5030 Da之间的毒素。大多数毒素(5)含有6个二硫键，1个毒素含有8个二硫键。我得到了几个新的毒液导管，从中提取了毒液，曼德用高效液相色谱分离了毒液，但数量正好在检测极限上。我保存了一小部分原始毒液，并将尝试使用原始材料对毒素进行排序。对于这些物种，绝对没有可用的数据库，我们拥有的材料数量也少得惊人。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. The aim of the project is to identify toxins from Terebridae and Turret species. The peptides present in the crude venom of Terebridae gutatta, argyosia, maunti and formosa were measured by LC-MS on the Orbitrap. Only the gutatta and argyosia venom had enough material to allow off-line separation. These two venoms were separated by RP-HPLC and the fractions exhibiting strong UV absorption at 214 nm were analyzed with MALDI TOF mass spectrometry to determine the accurate mass and number of disulfide bonds of potential toxins in these fractions. The Terebridae aergyosia venom showed hardly any material using UV detection, but Terebridae gutatta showed 5 major peaks. Unfortunately using a standard reduction protocol all of the gutatta peptides precipitated. The reduction protocol was optimized by increasing the amount of organic solvent (acetonitrile and methanol) and addition of GnHCL. Toxins with masses ranging from 3700 to 5030 Da were identified. The majority of toxins (5) contained 6 disulfide bonds and one toxin contained 8 disulfide bonds. I obtained several new venom ducts from which I extracted the venom and Mande separated the by HPLC, but the amounts are right on the detection limit. I saved a small aliquot of the crude venoms and will try to sequence the toxin using the crude material. There is absolutely no database available for these species and the amounts of material we have are vanishingly small.