The mitochondria-targeted derivative of the classical uncoupler of oxidative phosphorylation carbonyl cyanide m-chlorophenylhydrazone is an effective mitochondrial recoupler

Iaubasarova, Iliuza R; Khailova, Ljudmila S.; Firsov, Alexander M.; Grivennikova, Vera G.; Kirsanov, Roman S.; Коршунова, Г. А.; Kotova, Еlena А.; Antonenko, Yuri N.

doi:10.1371/journal.pone.0244499

Cited by 17 publications

(6 citation statements)

References 37 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This suggestion is supported by direct experiments on the interaction between the azido derivative of CCCP and mitochondrial proteins [ 15 ]. A recent study at our laboratory showed that the CCCP–triphenylphosphonium conjugate, which does not uncouple mitochondria, is able to block the uncoupling effect of CCCP [ 53 ]. The involvement of a protein in the uncoupling activity of CCCP is also evidenced by the strong inhibition of the CCCP effect on mitochondria by 6-ketocholestanol, which, on the contrary, can increase the CCCP-induced proton current in BLM due to an elevation of the membrane dipole potential [ 54 ].…”

Section: Interaction Between Protonophores and Mitochondrial Membrane...mentioning

confidence: 99%

Fifty Years of Research on Protonophores: Mitochondrial Uncoupling As a Basis for Therapeutic Action

Kotova

Antonenko

2022

Acta Naturae

Self Cite

View full text Add to dashboard Cite

Protonophores are compounds capable of electrogenic transport of protons across membranes. Protonophores have been intensively studied over the past 50 years owing to their ability to uncouple oxidation and phosphorylation in mitochondria and chloroplasts. The action mechanism of classical uncouplers, such as DNP and CCCP, in mitochondria is believed to be related to their protonophoric activity; i.e., their ability to transfer protons across the lipid part of the mitochondrial membrane. Given the recently revealed deviations in the correlation between the protonophoric activity of some uncouplers and their ability to stimulate mitochondrial respiration, this review addresses the involvement of some proteins of the inner mitochondrial membrane, such as the ATP/ADP antiporter, dicarboxylate carrier, and ATPase, in the uncoupling process. However, these deviations do not contradict the Mitchell theory but point to a more complex nature of the interaction of DNP, CCCP, and other uncouplers with mitochondrial membranes. Therefore, a detailed investigation of the action mechanism of uncouplers is required for a more successful pharmacological use, including their antibacterial, antiviral, anticancer, as well as cardio-, neuro-, and nephroprotective effects.

show abstract

Section: Interaction Between Protonophores and Mitochondrial Membrane...mentioning

confidence: 99%

Fifty Years of Research on Protonophores: Mitochondrial Uncoupling As a Basis for Therapeutic Action

Kotova

Antonenko

2022

Acta Naturae

Self Cite

View full text Add to dashboard Cite

show abstract

“…To analyze whether the observed mitochondrial accumulation was associated with impaired mitophagy, we monitored the ability of the neurons to clear impaired mitochondria in response to CCCP [27]. To optimize the mitophagy response window, neurons were treated with 10 µM CCCP in a time-dependent manner (24,48,72, and 96 hrs) followed by quantification of nuclei and the area covered by the mitochondrial marker TOM20 (Suppl.…”

Section: Parkin Deficiency Results In Impaired Mitophagymentioning

confidence: 99%

USP30 inhibition induces mitophagy and reduces oxidative stress in parkin-deficient and CCCP-stressed human iPSC-derived neurons

Okarmus

Agergaard

Stummann

et al. 2022

Preprint

View full text Add to dashboard Cite

Ubiquitination of mitochondrial proteins plays an important role in the cellular regulation of mitophagy. The E3 ubiquitin ligase parkin (encoded by PARK2) and the ubiquitin-specific protease 30 (USP30) have both been reported to regulate ubiquitination of outer mitochondrial proteins and thereby mitophagy. Loss of E3 ligase activity is thought to be pathogenic in both sporadic and inherited Parkinson's disease (PD), with loss-of-function mutations in PARK2 being the most frequent cause of autosomal recessive PD. The aim of the present study was to evaluate whether mitophagy induced by USP30 inhibition provides a functional rescue in isogenic human induced pluripotent stem cell-derived dopaminergic neurons with and without PARK2 knockout (KO). Our data show that healthy neurons responded to CCCP-induced mitochondrial damage by clearing the impaired mitochondria and that this process was accelerated by USP30 inhibition. Parkin-deficient neurons showed an impaired mitophagic response to CCCP challenge, although mitochondrial ubiquitination was enhanced. USP30 inhibition promoted mitophagy in PARK2 KO neurons, independently of whether left in basal conditions or treated with CCCP. In PARK2 KO, as in control neurons, USP30 inhibition balanced oxidative stress levels by reducing excessive production of reactive oxygen species. Interestingly, non-dopaminergic neurons, were the main driver of the beneficial effects of USP30 inhibition. Our findings demonstrate that USP30 inhibition is a promising approach to boost mitophagy and improve cellular health, also in parkin-deficient cells, and support the potential relevance of USP30 inhibitors as a novel therapeutic approach in diseases with a need to combat neuronal stress mediated by impaired mitochondria.

show abstract

“…This result proved the applicability of the calibration plot ( Figure 3 B and Figure 4 B) and led us to reject the hypothesis of an unequal extra- and intracellular pH formed upon ionophore treatment. To verify the performance of the indicator under conditions stimulating a physiologically mediated pH shift in the mitochondria (i.e., to record the pH dynamics in live cells), we treated the cells with CCCP (carbonyl cyanide m-chlorophenyl hydrazone), an uncoupler of the mitochondrial electron transport chain, which inhibits oxidative phosphorylation through the uncoupling of the proton gradient in the inner mitochondrial membrane [ 33 ]. CCCP molecules spontaneously diffuse throughout the plasma membrane and enter the mitochondria, leading to a slight acidification of their matrix, which is well-described in the literature [ 33 ].…”

Section: Resultsmentioning

confidence: 99%

A Single Fluorescent Protein-Based Indicator with a Time-Resolved Fluorescence Readout for Precise pH Measurements in the Alkaline Range

Simonyan

Protasova

Mamontova

et al. 2022

IJMS

View full text Add to dashboard Cite

The real-time monitoring of the intracellular pH in live cells with high precision represents an important methodological challenge. Although genetically encoded fluorescent indicators can be considered as a probe of choice for such measurements, they are hindered mostly by the inability to determine an absolute pH value and/or a narrow dynamic range of the signal, making them inefficient for recording the small pH changes that typically occur within cellular organelles. Here, we study the pH sensitivity of a green-fluorescence-protein (GFP)-based emitter (EGFP-Y145L/S205V) with the alkaline-shifted chromophore’s pKa and demonstrate that, in the pH range of 7.5–9.0, its fluorescence lifetime changes by a factor of ~3.5 in a quasi-linear manner in mammalian cells. Considering the relatively strong lifetime response in a narrow pH range, we proposed the mitochondria, which are known to have a weakly alkaline milieu, as a target for live-cell pH measurements. Using fluorescence lifetime imaging microscopy (FLIM) to visualize the HEK293T cells expressing mitochondrially targeted EGFP-Y145L/S205V, we succeeded in determining the absolute pH value of the mitochondria and recorded the ETC-uncoupler-stimulated pH shift with a precision of 0.1 unit. We thus show that a single GFP with alkaline-shifted pKa can act as a high-precision indicator that can be used in a specific pH range.

show abstract

The mitochondria-targeted derivative of the classical uncoupler of oxidative phosphorylation carbonyl cyanide m-chlorophenylhydrazone is an effective mitochondrial recoupler

Cited by 17 publications

References 37 publications

Fifty Years of Research on Protonophores: Mitochondrial Uncoupling As a Basis for Therapeutic Action

Fifty Years of Research on Protonophores: Mitochondrial Uncoupling As a Basis for Therapeutic Action

USP30 inhibition induces mitophagy and reduces oxidative stress in parkin-deficient and CCCP-stressed human iPSC-derived neurons

A Single Fluorescent Protein-Based Indicator with a Time-Resolved Fluorescence Readout for Precise pH Measurements in the Alkaline Range

Contact Info

Product

Resources

About