Role of membrane microdomains in compartmentation of cAMP signaling.
Role of membrane microdomains in compartmentation of cAMP signaling.
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DOI：
10.1371/journal.pone.0095835
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Harvey RD
中科院分区：
文献类型：
--
作者：
Agarwal SR;Yang PC;Rice M;Singer CA;Nikolaev VO;Lohse MJ;Clancy CE;Harvey RD
在空间上将 cAMP 的产生限制在离散的亚细胞位置，可以选择性地调节特定的功能反应。但 cAMP 信号传导的具体限制位置和方式尚不完全清楚。负责 cAMP 产生的不同受体和腺苷酸环化酶亚型在质膜的脂筏和非脂筏结构域之间分布不均匀。我们试图确定这些膜结构域在组织 HEK293 细胞中的 cAMP 反应中所起的作用。基于自由扩散 FRET 的生物传感器 Epac2-camps 用于测量全局 cAMP 反应，而针对脂筏 (Epac2-MyrPalm) 和非脂筏 (Epac2-CAAX) 结构域的探针版本用于监测质膜附近的局部 cAMP 产生。胆固醇消耗对脂筏的破坏选择性地改变了脂筏相关受体产生的 cAMP 反应。结果表明，与脂筏以及非脂筏结构域相关的受体可以促进整体 cAMP 反应。此外，发现非筏结构域中的基础 cAMP 活性显着较高。腺苷酸环化酶活性的药物抑制会降低 Epac2-CAAX 检测到的基础 cAMP 活性，但不会降低 Epac2-MyrPalm 或 Epac2-camps 检测到的基础 cAMP 活性，这一事实支持了这一点。 Epac2-CAAX 检测到的反应对腺苷酸环化酶活性的直接刺激也更敏感，但对磷酸二酯酶活性的抑制不太敏感。定量模型用于证明腺苷酸环化酶和磷酸二酯酶活性的差异是必要的，但不足以解释与质膜不同微域相关的 cAMP 的区室。
Spatially restricting cAMP production to discrete subcellular locations permits selective regulation of specific functional responses. But exactly where and how cAMP signaling is confined is not fully understood. Different receptors and adenylyl cyclase isoforms responsible for cAMP production are not uniformly distributed between lipid raft and non-lipid raft domains of the plasma membrane. We sought to determine the role that these membrane domains play in organizing cAMP responses in HEK293 cells. The freely diffusible FRET-based biosensor Epac2-camps was used to measure global cAMP responses, while versions of the probe targeted to lipid raft (Epac2-MyrPalm) and non-raft (Epac2-CAAX) domains were used to monitor local cAMP production near the plasma membrane. Disruption of lipid rafts by cholesterol depletion selectively altered cAMP responses produced by raft-associated receptors. The results indicate that receptors associated with lipid raft as well as non-lipid raft domains can contribute to global cAMP responses. In addition, basal cAMP activity was found to be significantly higher in non-raft domains. This was supported by the fact that pharmacologic inhibition of adenylyl cyclase activity reduced basal cAMP activity detected by Epac2-CAAX but not Epac2-MyrPalm or Epac2-camps. Responses detected by Epac2-CAAX were also more sensitive to direct stimulation of adenylyl cyclase activity, but less sensitive to inhibition of phosphodiesterase activity. Quantitative modeling was used to demonstrate that differences in adenylyl cyclase and phosphodiesterase activities are necessary but not sufficient to explain compartmentation of cAMP associated with different microdomains of the plasma membrane.
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影响因子：
7.8
作者：
Kenworthy, Anne K;Nichols, Benjamin J;Remmert, Catha L;Hendrix, Glenn M;Kumar, Mukesh;Zimmerberg, Joshua;Lippincott-Schwartz, Jennifer
通讯作者：
Lippincott-Schwartz, Jennifer
影响因子：
5
作者：
Harvey RD;Calaghan SC
通讯作者：
Calaghan SC
影响因子：
--
作者：
Depry, Charlene;Allen, Michael D.;Zhang, Jin
通讯作者：
Zhang, Jin
DOI：
10.1073/pnas.2336102100
发表时间：
2003-11-25
影响因子：
11.1
作者：
Kwik, J;Boyle, S;Edidin, M
通讯作者：
Edidin, M
影响因子：
--
作者：
DiPilato, Lisa M.;Zhang, Jin
通讯作者：
Zhang, Jin