Literature DB >> 10700148

A fluorescent indicator for visualizing cAMP-induced phosphorylation in vivo.

Y Nagai1, M Miyazaki, R Aoki, T Zama, S Inouye, K Hirose, M Iino, M Hagiwara.   

Abstract

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.

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Year:  2000        PMID： 10700148     DOI： 10.1038/73767

Source DB:  PubMed          Journal:  Nat Biotechnol        ISSN： 1087-0156            Impact factor:   54.908


  37 in total

1.  Galpha i3 binding to calnuc on Golgi membranes in living cells monitored by fluorescence resonance energy transfer of green fluorescent protein fusion proteins.

Authors:  T S Weiss; C E Chamberlain; T Takeda; P Lin; K M Hahn; M G Farquhar
Journal:  Proc Natl Acad Sci U S A       Date:  2001-12-18       Impact factor: 11.205

2.  Genetically encoded reporters of protein kinase A activity reveal impact of substrate tethering.

Authors:  J Zhang; Y Ma; S S Taylor; R Y Tsien
Journal:  Proc Natl Acad Sci U S A       Date:  2001-12-18       Impact factor: 11.205

3.  Genetically encoded fluorescent reporters of protein tyrosine kinase activities in living cells.

Authors:  A Y Ting; K H Kain; R L Klemke; R Y Tsien
Journal:  Proc Natl Acad Sci U S A       Date:  2001-12-18       Impact factor: 11.205

4.  Homo-FRET microscopy in living cells to measure monomer-dimer transition of GFP-tagged proteins.

Authors:  I Gautier; M Tramier; C Durieux; J Coppey; R B Pansu; J C Nicolas; K Kemnitz; M Coppey-Moisan
Journal:  Biophys J       Date:  2001-06       Impact factor: 4.033

5.  Analysis of protein kinase A activity in insulin-secreting cells using a cell-penetrating protein substrate and capillary electrophoresis.

Authors:  Femina Rauf; Yiding Huang; Thusitha P Muhandiramlage; Craig A Aspinwall
Journal:  Anal Bioanal Chem       Date:  2010-05-12       Impact factor: 4.142

6.  Tuning FlaSh: redesign of the dynamics, voltage range, and color of the genetically encoded optical sensor of membrane potential.

Authors:  Giovanna Guerrero; Micah S Siegel; Botond Roska; Eli Loots; Ehud Y Isacoff
Journal:  Biophys J       Date:  2002-12       Impact factor: 4.033

7.  Correcting confocal acquisition to optimize imaging of fluorescence resonance energy transfer by sensitized emission.

Authors:  Jacco van Rheenen; Michiel Langeslag; Kees Jalink
Journal:  Biophys J       Date:  2004-04       Impact factor: 4.033

Review 8.  Genetically encoded sensors for metabolites.

Authors:  Karen Deuschle; Marcus Fehr; Melanie Hilpert; Ida Lager; Sylvie Lalonde; Loren L Looger; Sakiko Okumoto; Jörgen Persson; Anja Schmidt; Wolf B Frommer
Journal:  Cytometry A       Date:  2005-03       Impact factor: 4.355

Review 9.  Visualization of growth signal transduction cascades in living cells with genetically encoded probes based on Förster resonance energy transfer.

Authors:  Kazuhiro Aoki; Etsuko Kiyokawa; Takeshi Nakamura; Michiyuki Matsuda
Journal:  Philos Trans R Soc Lond B Biol Sci       Date:  2008-06-27       Impact factor: 6.237

10.  Optical lock-in detection of FRET using synthetic and genetically encoded optical switches.

Authors:  Shu Mao; Richard K P Benninger; Yuling Yan; Chutima Petchprayoon; David Jackson; Christopher J Easley; David W Piston; Gerard Marriott
Journal:  Biophys J       Date:  2008-02-15       Impact factor: 4.033
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