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

被引：11

作者：

Zhang, Meiqi ^{[1
,2
]}

Yang, Bo ^{[1
,2
]}

Zhang, Jiayuan ^{[1
,2
,3
]}

Song, Yuxin ^{[1
,2
]}

Wang, Weibo ^{[1
,2
,4
]}

Li, Na ^{[1
,2
]}

Wang, Yuan ^{[1
,2
]}

Li, Wenzhe ^{[1
,2
]}

Wang, Jing ^{[1
,2
]}

机构：

[1] Peking Univ, State Key Lab Nat & Biomimet Drugs, Beijing 100191, Peoples R China

[2] Peking Univ, Dept Chem Biol, Sch Pharmaceut Sci, Beijing 100191, Peoples R China

[3] Univ Oxford, Wellcome Ctr Human Genet, Roosevelt Dr, Oxford OX3 7BN, England

[4] Cent China Normal Univ, Key Lab Pesticide & Chem Biol, Minist Educ,Coll Chem, Int Joint Res Ctr Intelligent Biosensor Technol &, Wuhan 430079, Peoples R China

来源：

ANGEWANDTE CHEMIE-INTERNATIONAL EDITION | 2022年 / 61卷 / 33期

基金：

中国国家自然科学基金;

关键词：

Biosensor; GTP; GDP Ratio; Imaging; Metabolism; Mitochondria; LIVE CELLS; GLUCOSE; ATP; NUCLEOTIDES; INHIBITION; EXPRESSION; MECHANISM; SENSORS; EIF5B; STATE;

D O I：

10.1002/anie.202201266

中图分类号：

O6 [化学];

学科分类号：

0703 ;

摘要：

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.

引用

页数：11

共 49 条

[1] Genetically encoded fluorescent indicator for intracellular hydrogen peroxide [J].