MEASURMENT OF DAG MOLECULAR SPECIES IN BASOPHILS

嗜碱性粒细胞中 DAG 分子种类的测量

• 批准号: 2023459
• 负责人: WALTER C HUBBARD
• 金额: $ 17.38万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-05-01 至 1998-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2023459
• 关键词: analytical method; basophils; diacylglycerols; gas chromatography mass spectrometry; technology /technique development

DESCRIPTION (Adapted from the Investigator's abstract): Diacylglycerols (DAG) have a central role in early receptor-mediated signal transduction events for many types of cells. These lipid molecules also appear to be involved in cell differentiation, cell growth and apoptosis. Previously existing assays for this class of molecules are sensitive only in the picomole range, if total DAG levels are examined, or require significantly higher levels if one is interested in a molecular species profile. There are many questions regarding the relative role of DAG species as well as questions about when and where they are generated. This application seeks to refine a new technique to measure the generation of diradylglycerols, acyl or alkyl linked. This technique provides 1000 fold better sensitivity than conventional methods, molecular species specificity and mass measurements of these species. The method employs gas chromatography/negative ion chemical ionization mass spectrometry (GC/NICIMS) to separate and measure selected molecular species of DAG using internal deuterated standards for quantitation. The application seeks to fully develop this approach so that both molecular species of DAG and phospholipids may be analyzed with high sensitivity. In addition, a new method to profile phospholipid species, without a prior knowledge of the species present is proposed. When this latter method is fully developed, it may be possible to predict which DAG species deserve attention using our other proposed methodologies. This latter technique should allow investigators to understand the extent to which cells selectively generate specific molecular species. We naturally expect that having a highly sensitive assay for DAGs will facilitate studies of this important second messenger in a wide variety of cells systems and cell organelles.

性状（改编自研究者摘要）：二酰基甘油 (DAG)在早期受体介导的信号转导中起中心作用 许多类型的细胞。 这些脂质分子似乎也是 参与细胞分化、细胞生长和凋亡。 先前 对这类分子的现有测定仅在 皮摩尔范围，如果检查总DAG水平，或需要显著 更高的水平，如果人们对分子物种谱感兴趣的话。 那里 关于DAG物种的相对作用以及 关于它们何时何地产生的问题。 本申请寻求 为了改进一种新技术来测量二氢甘油的产生， 酰基或烷基连接。 这项技术提供了1000倍的灵敏度 比传统方法，分子种类特异性和质量 对这些物种的测量。 该方法使用气体， 色谱/负离子化学电离质谱 (GC/NICIMS）分离和测量DAG的选定分子种类， 用于定量的氘代内标物。 该申请旨在 充分发展这种方法，使这两种分子物种的DAG和 可以高灵敏度分析磷脂。 一种新 方法来剖析磷脂种类，没有事先的知识， 目前的物种。 当后一种方法得到充分发展时， 也许可以预测哪些DAG物种值得关注，使用我们的 其他拟议的方法。 后一种技术应允许 研究人员了解细胞选择性产生的程度 特定的分子种类。 我们自然希望有一个高度 DAGs的灵敏测定将有助于这一重要的第二个研究 在多种细胞系统和细胞器中的信使。