Subcellular dynamics of protein kinase A activity visualized by FRET-based reporters
Subcellular dynamics of protein kinase A activity visualized by FRET-based reporters
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DOI：
10.1016/j.bbrc.2006.07.136
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发表时间：
2006-09-22
影响因子：
3.1
通讯作者：
Zhang, Jin
中科院分区：
文献类型：
--
作者：
Allen, Michael D.;Zhang, Jin
cAMP依赖性蛋白激酶（PKA）通过区室化活性介导关键的细胞过程，并且在活细胞中跟踪其活性的能力应该有助于增加我们对这种精确调节的理解。在这里，通过对新荧光蛋白的系统测试，我们开发了一种新的基于FRET的A激酶活性报告基因（AKAR）AKAR 3，其动态范围为31- 41%，是前人的两倍。在质膜，细胞质，细胞核和线粒体的PKA活性的可视化实现。将AKAR 3靶向到线粒体外膜揭示了线粒体处的基础PKA活性与细胞质中的PKA活性不同，表明PKA活性在不同亚细胞位置处的差异调节。(c)2006年爱思唯尔公司All rights reserved.
cAMP-dependent protein kinase (PKA) mediates key cellular processes via compartmentalized activity, and the ability to track its activity in living cells should help increase our understanding of this precise regulation. Here, through systematic testing of new fluorescent proteins, we developed a new FRET-based A-kinase activity reporter (AKAR), AKAR3, with a dynamic range of 31-41%, twice that of predecessors. Visualization of PKA activity at plasma membrane, cytoplasm, nucleus, and mitochondria was achieved. Targeting AKAR3 to outer mitochondrial membrane revealed that basal PKA activity at mitochondria differs from that in the cytoplasm, indicating differential regulation of PKA activity at different subcellular locations. (c) 2006 Elsevier Inc. All rights reserved.