Calcium Influx Factor

钙流入因子

• 批准号: 7903957
• 负责人: Victoria M Bolotina
• 金额: $ 21.13万
• 依托单位: BOSTON MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-08-14 至 2011-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7903957
• 关键词: Achievement; Arts; Attention; Binding; Biochemical Pathway; Biological Assay; Biological Testing; Blood Vessels; Calcium; Calmodulin; Cations; Cell membrane; Cells; Chemicals; Collaborations; Complex; Data; Disease; Endoplasmic Reticulum; Enzymes; Felis catus; Goals; Health; Homeostasis; Human; Knowledge; Laboratories; Life; Lysophospholipids; Mass Spectrum Analysis; Membrane; Methods; Molecular; Molecular Structure; Pathway interactions; Phospholipase A2; Physiological; Platelet Factor 4; Production; Recording of previous events; Research; Research Personnel; Role; STIM1 gene; Sampling; Second Messenger Systems; Signal Transduction; Source; Structure; System; Techniques; Testing; United States National Institutes of Health; Work; Yeasts; analog; cell type; experience; innovation; novel; novel strategies; programs; public health relevance; second messenger; sensor; tool

DESCRIPTION (provided by applicant): The long term goal of the PI's laboratory is to define the molecular components and mechanism of the store- operated Ca2+ entry (SOCE), which is known to be activated upon depletion of intracellular Ca2+ stores in excitable and non-excitable cells The focus of this innovative proposal is on calcium influx factor (CIF), a notoriously elusive and still un- identified messenger, that is produced upon depletion of the stores and was found in all living cells tested so far (from yeast to humans). We have unique knowledge and extensive experience in working with CIF extracts. We had discovered the physiological target for CIF (a specific plasma membrane bound Ca2+ independent phospholipase A2), and demonstrated its crucial role in SOCE, which was confirmed by other investigators. Most recently, we identified STIM1 as a trigger for CIF production in the endoplasmic reticulum. During our previous studies that focused on the mechanism of CIF production, action and its role in SOCE pathway, PI's laboratory had accumulated unique knowledge and expertise that may finally allow molecular identification of this extremely important but still elusive messenger. We had established new methods for advanced multi-step purification and biological testing of CIF, and can obtain sufficient quantities of CIF of the highest purity that is essential for its instrumental analysis. The goal of our new proposal is molecular identification of CIF. We understand that this is an ambitious goal, but we believe that we finally have everything for its successful achievement: 1) the knowledge about the molecular trigger of CIF production, and about physiological target of CIF, 2) extensive experience in working with CIF extracts, 3) new abundant source of CIF, 4) all the tools and novel approaches for its fine purification, 5) new effective and sensitive bioassay approaches and methods for testing samples and candidate molecules for CIF activity, 6) state-of-the-art facilities and established collaborations with the world class experts in mass spectrometry, NMR and SOCE mechanism. The feasibility of CIF purification and identification of its molecular structure is fully supported by our unique expertise, and extensive preliminary data. Specific Aim of this proposal is to identify CIF. We will: 1.1. Determine chemical and structural components of CIF using advanced mass spectrometry and NMR techniques. 1.2. Synthesize CIF molecule and its inactive analogs. 1.3. Verify CIF identity in bioassay systems, and match it with endogenous CIF activity. 1.4. Identify the biochemical pathway for CIF production. PUBLIC HEALTH RELEVANCE: The goal of this proposal is to identify calcium influx factor (CIF), a novel second messenger that is ubiquitously produced by all the cell types tested so far (from yeast to human), and is involved in activation of the store-operated Ca2+ entry pathway, one of the major mechanisms that determine Ca2+ homeostasis in health and disease. The feasibility of CIF identification is fully supported by our unique expertise, extensive preliminary data and advanced approaches that are already developed in PI's lab.

描述（由申请人提供）：PI 实验室的长期目标是定义钙库操作的 Ca2+ 进入 (SOCE) 的分子成分和机制，已知该通道在兴奋性和非兴奋性细胞内 Ca2+ 储存耗尽时被激活。 -兴奋细胞 这项创新提案的重点是钙流入因子（CIF），这是一种众所周知的难以捉摸且仍未被识别的信使，它是在储存耗尽时产生的，并且迄今为止测试的所有活细胞（从酵母到人类）中都发现了这种物质。我们在处理 CIF 提取物方面拥有独特的知识和丰富的经验。我们发现了 CIF（一种特定的质膜结合 Ca2+ 独立磷脂酶 A2）的生理靶点，并证明了它在 SOCE 中的关键作用，这一点得到了其他研究人员的证实。最近，我们发现 STIM1 是内质网中 CIF 产生的触发因素。在我们之前的研究中，重点关注 CIF 的产生、作用及其在 SOCE 途径中的作用机制，PI 实验室积累了独特的知识和专业知识，最终可能能够对这一极其重要但仍然难以捉摸的信使进行分子鉴定。我们建立了 CIF 先进多步纯化和生物测试的新方法，可以获得足够数量的最高纯度的 CIF，这对于仪器分析至关重要。我们新提案的目标是 CIF 的分子鉴定。我们知道这是一个雄心勃勃的目标，但我们相信我们最终拥有成功实现这一目标所需的一切：1）有关 CIF 产生的分子触发因素以及 CIF 生理目标的知识，2）在使用 CIF 提取物方面拥有丰富的经验，3）新的丰富的 CIF 来源，4）用于其精细纯化的所有工具和新颖方法，5）用于测试样品和候选分子的 CIF 活性的新的有效且灵敏的生物测定方法和方法，6）最新状态艺术设施和与质谱、核磁共振和SOCE机制方面的世界级专家建立了合作。 CIF 纯化及其分子结构鉴定的可行性得到了我们独特的专业知识和广泛的初步数据的充分支持。该提案的具体目标是确定 CIF。我们将： 1.1。使用先进的质谱和 NMR 技术确定 CIF 的化学和结构成分。 1.2.合成 CIF 分子及其无活性类似物。 1.3.验证生物测定系统中的 CIF 身份，并将其与内源 CIF 活性进行匹配。 1.4.确定 CIF 生产的生化途径。公共健康相关性：该提案的目标是确定钙流入因子（CIF），这是一种新型第二信使，迄今为止测试的所有细胞类型（从酵母到人类）普遍产生，并参与钙库的激活-操纵的Ca2+进入途径，决定健康和疾病中Ca2+稳态的主要机制之一。 CIF 鉴定的可行性得到了我们独特的专业知识、广泛的初步数据和 PI 实验室已开发的先进方法的充分支持。