REV, a BRET-based sensor of ERK activity

Frontiers in Endocrinology

Laurent eFagni¹  Ignacio eVillamil¹  nathalie ebouquier¹  Julie ePerroy¹  Chanjuan eXu²  Jianfeng eLiu²  vincent eollendorff³  Marion ePeter⁴ 

[1] Centre National de la Recherche Scientifique;Huazhong University of Science and Technology;INRA;Institut de Génétique Moléculaire de Montpellier (IGMM);

关键词: biosensor;    Bioluminescence resonance energy transfer (BRET);    BRET imaging;    Fluorescence-lifetime imaging microscopy (FLIM);    Extracellular signal-regulated kinases (ERK);    spatio-temporal signaling;   

DOI  :  10.3389/fendo.2013.00095

来源: DOAJ

【 摘 要 】

Networks of signaling molecules are activated in response to environmental changes. How are these signaling networks dynamically integrated in space and time to process particular information? To tackle this issue, biosensors of single signaling pathways have been engineered. Bioluminescence resonance energy transfer (BRET)-based biosensors have proven to be particularly efficient in that matter due to the high sensitivity of this technology to monitor protein-protein interactions or conformational changes in living cells. Extracellular signal-regulated kinases (ERK) are ubiquitously expressed and involved in many diverse cellular functions that might be encoded by the strength and spatio-temporal pattern of ERK activation. We developed a BRET-based sensor of ERK activity, called « REV » for Rluc8-ERKsubstrate-Venus. As expected, BRET changes of REV were correlated with ERK phosphorylation, which is required for its kinase activity. In neurons, the nature of the stimuli determines the strength, the location or the moment of ERK activation, thus highlighting how acute modulation of ERK may encode the nature of initial stimulus to specify the consequences of this activation. This study provides evidence for suitability of REV as a new biosensor to address biological questions.

【 授权许可】

Unknown