Real-time imaging of the intracellular glutathione redox potential

被引：559

作者：

Gutscher, Marcus ^{[1
]}

Pauleau, Anne-Laure ^{[1
]}

Marty, Laurent ^{[2
]}

Brach, Thorsten ^{[2
]}

Wabnitz, Guido H. ^{[3
]}

Samstag, Yvonne ^{[3
]}

Meyer, Andreas J. ^{[2
]}

Dick, Tobias P. ^{[1
]}

机构：

[1] German Canc Res Ctr, Redox Regulat Res Grp, DKFZ A160, D-69120 Heidelberg, Germany

[2] Univ Heidelberg, Heidelberg Inst Plant Sci, D-69120 Heidelberg, Germany

[3] Univ Heidelberg, Inst Immunol, D-69120 Heidelberg, Germany

来源：

NATURE METHODS | 2008年 / 5卷 / 06期

关键词：

D O I：

10.1038/NMETH.1212

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Dynamic analysis of redox-based processes in living cells is now restricted by the lack of appropriate redox biosensors. Conventional redox-sensitive GFPs (roGFPs) are limited by undefined specificity and slow response to changes in redox potential. In this study we demonstrate that the fusion of human glutaredoxin-1 (Grx1) to roGFP2 facilitates specific real-time equilibration between the sensor protein and the glutathione redox couple. The Grx1-roGFP2 fusion protein allowed dynamic live imaging of the glutathione redox potential (E-GSH) in different cellular compartments with high sensitivity and temporal resolution. The biosensor detected nanomolar changes in oxidized glutathione (GSSG) against a backdrop of millimolar reduced glutathione (GSH) on a scale of seconds to minutes. It facilitated the observation of redox changes associated with growth factor availability, cell density, mitochondrial depolarization, respiratory burst activity and immune receptor stimulation.

引用

页码：553 / 559

页数：7

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