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1 March 2019 G protein-assisted optimization of GPCR-activation based (GRAB) sensors
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Proceedings Volume 10865, Neural Imaging and Sensing 2019; 108650N (2019) https://doi.org/10.1117/12.2514631
Event: SPIE BiOS, 2019, San Francisco, California, United States
Abstract
In the brain, neurotransmitters or neuromodulators play pivotal roles in chemical synaptic transmission and consequently, monitoring their dynamics, especially in vivo, is critical for understanding their physiological- or pathophysiological roles at molecular, cellular, and circuit levels during behaviors and/or during diseases. We recently developed genetically-encoded GPCR-activation based (GRAB) sensors capable of reporting dynamics of acetylcholine, dopamine and norepinephrine with rapid kinetics, chemical- and cell-specificity in multiple organisms in vivo. Here, we explored the usage of G protein derivatives, either mini-G proteins or C-terminal peptides of Gα subunit to engineer new GRAB sensors. We found that the conformational changes mediated by mini-G proteins interacting with GPCRs, or Gα Cterminal peptides interacting with GPCRs could be harnessed to regulate fluorescence outputs of a GPCR fused circular permuted GFP (cpGFP). In addition, inter-molecular fusion of Gα C-terminal peptides significantly suppressed ectopic activation of G protein signaling in a GRAB acetylcholine sensor. Finally, we showed Gα C-terminal peptides fusion strategy could be applied to generate various GRAB sensors for small molecular compounds or neuropeptides.
Conference Presentation
© (2019) COPYRIGHT Society of Photo-Optical Instrumentation Engineers (SPIE). Downloading of the abstract is permitted for personal use only.
Zhaofa Wu, Jiesi Feng, Miao Jing, and Yulong Li "G protein-assisted optimization of GPCR-activation based (GRAB) sensors", Proc. SPIE 10865, Neural Imaging and Sensing 2019, 108650N (1 March 2019); https://doi.org/10.1117/12.2514631
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