CHARACTERIZING TEREBRIDAE TOXINS

拟肢虫毒素的特征

• 批准号: 8361560
• 负责人: Mande N. Holford
• 金额: $ 0.78万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-03-01 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8361560
• 关键词: Acetonitriles; Aliquot; Databases; Detection; Duct (organ) structure; Exhibits; Funding; Grant; High Pressure Liquid Chromatography; MALDI-TOF Mass Spectrometry; Measures; Methanol; National Center for Research Resources; Organic solvent product; Peptides; Principal Investigator; Protocols documentation; Research; Research Infrastructure; Resources; Right-On; Source; Taiwan; Toxin; United States National Institutes of Health; Venoms; absorption; cost; disulfide bond; macromolecule

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.

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 该项目的目的是确定Terebridae和Turret物种的毒素。本文采用液相色谱-质谱联用技术，在Orbitrap上测定了四种斑胸圆螺（Terebridae gutatta）、银斑圆螺（argyosia）、曼氏圆螺（Maunti）和福尔摩沙（formosa）粗毒中的多肽。只有gutta和argyosia毒液有足够的材料允许离线分离。 通过RP-HPLC分离这两种毒液，并使用MALDI TOF质谱法分析在214 nm处表现出强UV吸收的级分，以确定这些级分中潜在毒素的二硫键的准确质量和数量。 用紫外检测法，光刺螺毒液几乎没有显示任何物质，但光刺螺毒液显示5个主峰。不幸的是，使用标准还原方案，所有的古塔肽沉淀。通过增加有机溶剂（乙腈和甲醇）的量和添加GnHCL来优化还原方案。确定了质量范围为3700至5030 Da的毒素。大多数毒素（5种）含有6个二硫键，1种毒素含有8个二硫键。我得到了几个新的毒液管道，从中我提取了毒液，曼德用高效液相色谱法分离了毒液，但数量正好在检测限上。我留了一小部分粗毒液试着用粗毒液来测定毒素的序列。绝对没有这些物种的数据库，我们拥有的材料数量少得可怜。