Komlodi 2022 Abstract Bioblast
|3.1. «10+5» https://doi.org/10.26124/bec:2022-0001 |
Event: Bioblast 2022
The protonmotive force pmF establishes the link between electrical and chemical components of energy transformation and coupling in oxidative phosphorylation in the mitochondrial electron transfer system. The electrical part is corresponding to the mitochondrial membrane potential ΔΨmt and the chemical part is related to the transmembrane pH gradient ΔpH. Although the contribution of ΔpH to pmF is smaller than that of ΔΨmt, ΔpH plays an important role in mitochondrial transport processes and regulation of reactive oxygen species production. Separate measurement of ΔΨmt and ΔpH allows for calculation of pmF. Methods for monitoring ΔΨmt such as fluorescence dyes are generally available, while determination of ΔpH is more challenging.
In this review, we focus on the application of the fluorescence ratiometric method using the acetoxymethyl ester form of 2,7-biscarboxyethyl-5(6)-carboxyfluorescein (BCECF/AM) for real-time monitoring of the intramitochondrial pH in isolated mitochondria. Knowing the intra- and extramitochondrial pH allows for calculating the ΔpH. Application of specific ionophores such as nigericin or valinomycin, exerts the possibility to dissect the two components of the pmF in different directions. Furthermore, we tried to summarize those mitochondrial processes, such as production of reactive oxygen species, where the ΔpH has an important role.
• Keywords: BCECF; intramitochondrial pH; matrix pH; ΔpH; mitochondria, mitochondrial membrane potential, ΔΨmt; nigercin; protonmotive force, pmF; reverse electron transfer, RET; safranin; triphenylphosphonium, TPP+; valinomycin
• O2k-Network Lab: HU Budapest Tretter L
- Department of Biochemistry, Semmelweis University, Budapest, Hungary
List of abbreviations, terms and definitions - MitoPedia
Labels: MiParea: Instruments;methods, mt-Membrane
Preparation: Isolated mitochondria Enzyme: Complex V;ATP synthase Regulation: ATP production, mt-Membrane potential, pH Coupling state: LEAK, OXPHOS
HRR: Oxygraph-2k, O2k-Fluorometer, TPP Event: A3