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

Autor: Zhang M; State Key Laboratory of Natural and Biomimetic Drugs and Department of Chemical Biology, Department of Chemical Biology, School of Pharmaceutical Sciences, Peking University, Peking University, Beijing, 100191, China., Yang B; State Key Laboratory of Natural and Biomimetic Drugs and Department of Chemical Biology, Department of Chemical Biology, School of Pharmaceutical Sciences, Peking University, Peking University, Beijing, 100191, China., Zhang J; State Key Laboratory of Natural and Biomimetic Drugs and Department of Chemical Biology, Department of Chemical Biology, School of Pharmaceutical Sciences, Peking University, Peking University, Beijing, 100191, China.; Wellcome Centre for Human Genetics, University of Oxford, Roosevelt Dr, Headington, Oxford, OX3 7BN, UK., Song Y; State Key Laboratory of Natural and Biomimetic Drugs and Department of Chemical Biology, Department of Chemical Biology, School of Pharmaceutical Sciences, Peking University, Peking University, Beijing, 100191, China., Wang W; State Key Laboratory of Natural and Biomimetic Drugs and Department of Chemical Biology, Department of Chemical Biology, School of Pharmaceutical Sciences, Peking University, Peking University, Beijing, 100191, China.; Key Laboratory of Pesticide & Chemical Biology of Ministry of Education, International Joint Research Center for Intelligent Biosensor Technology and Health, College of Chemistry, Central China Normal University, Wuhan, 430079, China., Li N; State Key Laboratory of Natural and Biomimetic Drugs and Department of Chemical Biology, Department of Chemical Biology, School of Pharmaceutical Sciences, Peking University, Peking University, Beijing, 100191, China., Wang Y; State Key Laboratory of Natural and Biomimetic Drugs and Department of Chemical Biology, Department of Chemical Biology, School of Pharmaceutical Sciences, Peking University, Peking University, Beijing, 100191, China., Li W; State Key Laboratory of Natural and Biomimetic Drugs and Department of Chemical Biology, Department of Chemical Biology, School of Pharmaceutical Sciences, Peking University, Peking University, Beijing, 100191, China., Wang J; State Key Laboratory of Natural and Biomimetic Drugs and Department of Chemical Biology, Department of Chemical Biology, School of Pharmaceutical Sciences, Peking University, Peking University, Beijing, 100191, China.
Jazyk: angličtina
Zdroj: Angewandte Chemie (International ed. in English) [Angew Chem Int Ed Engl] 2022 Aug 15; Vol. 61 (33), pp. e202201266. Date of Electronic Publication: 2022 Jul 08.
DOI: 10.1002/anie.202201266
Abstrakt: 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.
(© 2022 Wiley-VCH GmbH.)
Databáze: MEDLINE
Externí odkaz: