Monitoring the Dynamic Regulation of the Mitochondrial GTP-to-GDP Ratio with a Genetically Encoded Fluorescent Biosensor

作者：Zhang, Meiqi; Yang, Bo; Zhang, Jiayuan; Song, Yuxin; Wang, Weibo; Li, Na; Wang, Yuan; Li, Wenzhe; Wang, Jing^*

来源：Angewandte Chemie - International Edition, 2022, 61(33): e202201266.

DOI：10.1002/anie.202201266

• Biosensor
• GTP
• GDP Ratio
• Imaging
• Metabolism
• Mitochondria

The interconversion of guanosine triphosphate (GTP) and guanosine diphosphate (GDP) is known to be integral to a wide variety of biological cellular activities, yet to date there are no analytical methods available to directly detect the ratio of intracellular GTP to GDP. Herein, we report GRISerHR, a genetically encoded fluorescent biosensor to monitor the GTP : GDP ratio in multiple cell types and in various organelles under metabolic perturbation. Additionally, we characterized the differential mitochondrial GTP : GDP ratios resulting from genetic modulation of two isoforms of a tricarboxylic acid (TCA) cycle enzyme (succinyl-CoA synthetase; SCS-ATP and SCS-GTP) and of a phosphoenolpyruvate (PEP) cycle enzyme (PEPCK-M). Thus, our GRISerHR sensor achieves spatiotemporally precise detection of dynamic changes in the endogenous GTP : GDP ratio in living cells and can help deepen our understanding about the energy metabolic contributions of guanosine nucleotides in biology.

• 单位
  北京大学