New Approach to Measuring ATP in Diabetes

摘要

Mitochondrial function is commonly measured by quantification ATP production using fluorescence or chemiluminescence assays and also by ~(31)P NMR spectrometry. In this issue of Diabetes (p. 1833), Yu et al. describe a new, highly sensitive, high-throughput ATP assay to measure mitochondrial function. The new assay relies on conversion of 2-deoxyglucose (2DOG) to 2-deoxyglucose phosphate (2D0GP) using the phosphate from ATP, thereby resulting in ADP formation. The extent of 2D0G to 2D0GP conversion depends on ATP produced by the mitochondria. Therefore, mitochondrial respiration or mitochondrial ATP production is measured as a function of 2D0GP formation. Because 2DOG to 2DOGP conversion is rapid and irreversible, the assay can measure respiration as a function of membrane potential at clamped ADP levels. The assay was validated by demonstrating that mitochondria isolated from diabetic muscle produces less 2DOGP (or ATP) compared with control muscle.