INTRACELLULAR CALCIUM CONTROL OF cAMP SYNTHESIS

坎普合成的细胞内钙控制

• 批准号: 6499346
• 负责人: DERMOT M COOPER
• 金额: $ 35.98万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-08-01 至 2005-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6499346
• 关键词: adenylate cyclase; bioengineering /biomedical engineering; biological signal transduction; biosensor device; calcium; calcium binding protein; calcium flux; calcium indicator; cell line; chemical kinetics; chimeric proteins; cyclic AMP; enzyme induction /repression; fluorescent dye /probe; luciferin monooxygenase; measurement; membrane channels; olfactions; phosphodiesterases; protein localization; protein structure function; second messengers; single cell analysis; site directed mutagenesis; technology /technique development; voltage /patch clamp

DESCRIPTION (Adapted from applicant's description): Adenylyl cyclases generate the ubiquitous second messenger cAMP. The importance of this messenger in regulating a large number of cellular processes is acknowledged. However, the methods for measuring cAMP lack both temporal and spatial resolution, leading to the widespread belief that these signals are simple. Against this notion, the large number of interactions between signaling pathways for Ca2+ and cAMP have been demonstrated and the dynamic nature of Ca2+ makes it likely that cAMP signals are also complex. In particular, Ca2+-sensitive adenylyl cyclases are regulated discreetly by physiological modes of Ca2+ entry and cAMP modulates Ca2+ entry by a variety of mechanisms. The present application proposes to develop adenylyl cyclase/ aequorin and adenylyl cyclase/ cameleon chimeras as localized sensors of Ca2+ and to develop olfactory cyclic nucleotide gated (oCNG) channels as rapid membrane-bound sensors of cAMP. Once these sensors are optimized, the applicant intends to examine dynamic and interdependent changes in the two signals in pituitary-derived GH4 cells, an excitable cell type. He will also use the cAMP sensor to study the rapid kinetics of adenylyl cyclase regulation in its native environment. These studies represent a first attempt to associate cAMP and Ca2+ signaling on the same temporal and spatial scale. In the long-term, such information will lead to greater generalized understanding of how the coordinated activity of these two second messengers control cellular function.

描述（改编自申请人的描述）：腺苷酸环化酶产生 普遍存在的第二信使cAMP。这位信使的重要性 调节大量细胞过程是公认的。但 测量cAMP的方法缺乏时间和空间分辨率，导致 人们普遍认为这些信号很简单。与此相反， Ca 2+和cAMP信号通路之间的大量相互作用 已经被证明，Ca 2+的动态性质使得cAMP可能 信号也很复杂。特别地，Ca 2+敏感的腺苷酸环化酶是 通过Ca 2+进入和cAMP调节的生理模式谨慎地调节 Ca 2+通过多种机制进入。本申请提出 开发腺苷酸环化酶/水母发光蛋白和腺苷酸环化酶/ cameleon嵌合体， 局部化的Ca 2+传感器，并开发嗅觉环核苷酸门控 （oCNG）通道作为cAMP的快速膜结合传感器。一旦这些传感器 优化，申请人打算检查动态和相互依存的变化 在垂体衍生的GH4细胞的两个信号中，一种可兴奋的细胞类型。他 还将使用cAMP传感器研究腺苷酸环化酶的快速动力学 在其自然环境中进行调控。这些研究代表了 在相同的时间和空间尺度上关联cAMP和Ca 2+信号传导。在 从长远来看，这些信息将导致更广泛的理解 这两种第二信使的协调活动如何控制细胞 功能