Chinopoulos 2009

Chinopoulos C, Vajda S, Csanady L, Mandi M, Mathe K, Adam-Vizi V (2009) A Novel Kinetic Assay of Mitochondrial ATP-ADP Exchange Rate Mediated by the ANT. Biophys J 96, 2490-504.

» PMID: 24391134

Chinopoulos Christos, Vajda Szilvia, Csanady Laszlo, Mandi Miklos, Mathe Katalin, Adam-Vizi Vera (2009) Biophys J

Abstract: A novel method exploiting the differential affinity of ADP and ATP to Mg(2+) was developed to measure mitochondrial ADP-ATP exchange rate. The rate of ATP appearing in the medium after addition of ADP to energized mitochondria, is calculated from the measured rate of change in free extramitochondrial [Mg²⁺] reported by the membrane-impermeable 5K⁺ salt of the Mg²⁺-sensitive fluorescent indicator, Magnesium Green, using standard binding equations. The assay is designed such that the adenine nucleotide translocase (ANT) is the sole mediator of changes in [Mg²⁺] in the extramitochondrial volume, as a result of ADP-ATP exchange. We also provide data on the dependence of ATP efflux rate within the 6.8-7.8 matrix pH range as a function of membrane potential. Finally, by comparing the ATP-ADP steady-state exchange rate to the amount of the ANT in rat brain synaptic, brain nonsynaptic, heart and liver mitochondria, we provide molecular turnover numbers for the known ANT isotypes. • Keywords: Adenine nucleotide translocase, ANT, ATP-ADP exchange, Magnesium Green • Bioblast editor: Cardoso Luiza HD,
• O2k-Network Lab: HU Budapest Chinopoulos C

Cited by

• Cardoso et al (2021) Magnesium Green for fluorometric measurement of ATP production does not interfere with mitochondrial respiration. Bioenerg Commun 2021.1. doi:10.26124/bec:2021-0001

Labels:

Organism: Rat  Tissue;cell: Heart, Nervous system, Liver  Preparation: Isolated mitochondria  Enzyme: Adenine nucleotide translocase  Regulation: ADP, ATP, ATP production 

MgG, Safranin, ANT, MitoFit 2021 MgG 

» Fluorometry protocol for Magnesium green