Monitoring the Dynamic Regulation of the Mitochondrial GTP-to-GDP Ratio with a Genetically Encoded Fluorescent Biosensor
Monitoring the Dynamic Regulation of the Mitochondrial GTP-to-GDP Ratio with a Genetically Encoded Fluorescent Biosensor
复制标题
使用基因编码荧光生物传感器监测线粒体 GTP 与 GDP 比率的动态调节
DOI：
10.1002/anie.202201266
复制
发表时间：
2022-07-08
影响因子：
16.6
通讯作者：
Wang，Jing
中科院分区：
文献类型：
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作者：
Zhang，Meiqi;Yang，Bo;Wang，Jing
已知鸟苷三磷酸（GTP）和鸟苷二磷酸（GDP）的相互转化是多种生物细胞活性的组成部分，但迄今为止还没有可用于直接检测细胞内GTP与GDP的比率的分析方法。在此，我们报告GRISerHR，基因编码的荧光生物传感器监测GTP：GDP的比例在多种细胞类型和各种细胞器下的代谢扰动。此外，我们还表征了由三羧酸（TCA）循环酶（琥珀酰辅酶A合成酶; SCS-ATP和SCS-GTP）和磷酸烯醇丙酮酸（PEP）循环酶（PEPCK-M）的两种亚型的遗传调节引起的线粒体GTP：GDP比值差异。因此，我们的GRISerHR传感器实现了对活细胞中内源性GTP：GDP比率动态变化的时空精确检测，并有助于加深我们对鸟苷核苷酸在生物学中的能量代谢贡献的理解。
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.