CHARACTERIZING TEREBRIDAE TOXINS

拟肢虫毒素的特征

• 批准号: 8169189
• 负责人: Mande N. Holford
• 金额: $ 0.35万
• 依托单位: ROCKEFELLER UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-03-01 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8169189
• 关键词: Acetonitriles; Computer Retrieval of Information on Scientific Projects Database; Data; Detection; Exhibits; Funding; Grant; High Pressure Liquid Chromatography; Institution; MALDI-TOF Mass Spectrometry; Measures; Methanol; Organic solvent product; Peptides; Protocols documentation; Research; Research Personnel; Resources; Solutions; Source; Taiwan; Tarantula Venoms; Time; Toxin; United States National Institutes of Health; Venoms; Work; absorption; aqueous; disulfide bond; method development

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. 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). Using high organic solvent the toxins remained in solution upon reduction. 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. To date, we are not able to obtain fragmentation data of these toxins as we have not yet determined conditions that keep the toxins in solution and allow analysis with RP HPLC. We are currently working out protocols that keep these hydrophobic toxins in solution and at the same time retained on RP-HPLC columns. This method development is also necessary for analyzing tarantula venoms that also precipitate out of aqueous solution upon reduction.

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 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）的量来优化还原方案。使用高有机溶剂，毒素在还原时保留在溶液中。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。迄今为止，我们无法获得这些毒素的碎片数据，因为我们尚未确定将毒素保持在溶液中并允许使用RP HPLC进行分析的条件。我们目前正在制定方案，使这些疏水性毒素保持在溶液中，同时保留在RP-HPLC柱上。这种方法的发展也是必要的分析狼蛛毒液，也沉淀出水溶液还原。