Calcium Influx Factor

钙流入因子

• 批准号: 7752223
• 负责人: Victoria M Bolotina
• 金额: $ 25.28万
• 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/7752223
• 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.

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在商店耗竭后产生，并在迄今已测试的所有活细胞中发现（从酵母到人类）。我们在使用CIF提取物方面拥有独特的知识和丰富的经验。我们已经发现了CIF的生理靶标（特定的质膜结合Ca2+独立的磷脂酶A2），并证明了其在SOCE中的关键作用，这是其他研究者证实的。最近，我们将STIM1确定为内质网中CIF产生的触发因素。在我们以前的研究中，PI的实验室积累了独特的知识和专业知识，该研究集中在CIF生产，行动及其在SOCE途径中的作用，最终可能允许分子鉴定这一极为重要但仍然难以捉摸的使者。我们已经建立了用于CIF的高级多步纯化和生物学测试的新方法，并可以获得足够数量的CIF的最高纯度，这对于其工具分析至关重要。我们新建议的目的是CIF的分子鉴定。 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对于CIF活动，6）最先进的设施，并与质谱，NMR和SOCE机制方面的世界一流专家建立了合作。我们独特的专业知识和广泛的初步数据完全支持CIF纯化和鉴定其分子结构的可行性。该建议的具体目的是识别CIF。我们将：1.1。使用晚期质谱和NMR技术确定CIF的化学和结构成分。 1.2。合成CIF分子及其非活性类似物。 1.3。验证生物测定系统中的CIF身份，并与内源性CIF活性匹配。 1.4。确定CIF生产的生化途径。公共卫生相关性：该提案的目的是鉴定钙涌入因子（CIF），这是一种新型的第二信使，迄今已测试的所有细胞类型（从酵母到人）都会生产，并且参与了商店经营的CA2+进入途径的激活，这是确定CA2+稳态的主要机制之一，这些机制之一确定了健康和疾病中的CA2+稳态。 CIF识别的可行性得到了我们独特的专业知识，广泛的初步数据以及PI实验室中已经开发的高级方法的充分支持。