A fluorescent indicator for visualizing cAMP-induced phosphorylation in vivo

被引：156

作者：

Nagai, Y

Miyazaki, M

Aoki, R

Zama, T

Inouye, S

Hirose, K

Iino, M

Hagiwara, M ^{[1
]}

机构：

[1] Tokyo Med & Dent Univ, Med Res Inst, Dept Funct Genom, Tokyo 1138510, Japan

[2] Univ Calif San Diego, Scripps Inst Oceanog, La Jolla, CA 92093 USA

[3] Univ Tokyo, Fac Med, Dept Pharmacol, Tokyo 1130033, Japan

来源：

NATURE BIOTECHNOLOGY | 2000年 / 18卷 / 03期

关键词：

phosphorylation; FRET; green fluorescent protein; imaging; PKA;

D O I：

10.1038/73767

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

We have developed a method for visualizing phosphorylation of proteins in living cells using a novel fluorescent indicator composed of two green fluorescent protein (GFP) variants joined by the kinase-inducible domain (KID) of the transcription factor cyclic adenosine monophosphate (cAMP)-responsive element binding protein (CREB). Phosphorylation of KID by the cAMP-dependent protein kinase A (PKA) decreased the fluorescence resonance energy transfer (FRET) among the flanking GFPs, By transfecting COS-7 cells with an expression vector encoding this indicator protein (termed ART for cAMP-responsive tracer), we were able to visualize activation dynamics of PKA in living cells.

引用

页码：313 / 316

页数：4

共 21 条

[1] FLUORESCENCE RATIO IMAGING OF CYCLIC-AMP IN SINGLE CELLS [J].