CNRS, UMR-5203, Institut de Génomique Fonctionnelle , Montpellier , France ; INSERM, U661 , Montpellier , France ; UMR-5203, Universités de Montpellier 1 & 2 , Montpellier , France ; Sino-France Laboratory for Drug Screening, Key Laboratory of Molecular Biophysics of Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology , Wuhan , China.
Front Endocrinol (Lausanne). 2013 Jul 30;4:95. doi: 10.3389/fendo.2013.00095. eCollection 2013.
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 Rluc8-ERKsubstrate-Venus (REV). 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.
信号分子网络会响应环境变化而被激活。这些信号网络如何在空间和时间上动态整合以处理特定信息？为了解决这个问题，人们已经设计出了用于检测单一信号通路的生物传感器。生物发光共振能量转移（BRET）生物传感器在这方面被证明是特别有效的，因为该技术具有很高的灵敏度，可以监测活细胞中的蛋白质-蛋白质相互作用或构象变化。细胞外信号调节激酶（ERK）广泛表达，并参与许多不同的细胞功能，这些功能可能由 ERK 激活的强度和时空模式来编码。我们开发了一种基于 BRET 的 ERK 活性传感器，称为 Rluc8-ERKsubstrate-Venus（REV）。正如预期的那样，REV 的 BRET 变化与 ERK 磷酸化相关，而磷酸化是其激酶活性所必需的。在神经元中，刺激的性质决定了 ERK 激活的强度、位置或时间，从而突出了 ERK 的急性调节如何可以编码初始刺激的性质，从而指定这种激活的后果。这项研究为 REV 作为一种新的生物传感器来解决生物学问题提供了证据。
