A Multi‐responsive Fluorescent Probe Reveals Mitochondrial Nucleoprotein Dynamics with Reactive Oxygen Species Regulation through Super‐resolution Imaging

Yang, Guanqing; Liu, Zhengjie; Zhang, Ruilong; Tian, Xiaohe; Chen, Juan; Liu, Bianhua; Han, Xinya; Fu, Yao; Hu, Zhangjun; Zhang, Zhongping

doi:10.1002/ange.202005959

Cited by 7 publications

(2 citation statements)

References 48 publications

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“…However, compounds 1 and 11 up to 100 μM displayed no significant toxicity with 95% cell survival, while entacapone without liver toxicity served as a negative control. The cell viability and mitochondria membrane potential is an indicator of cell viability 26 . Therefore, these data supported the idea that these novel non-nitrocatechol COMT inhibitors had the potential of being developed with a reduced risk of liver toxicity.…”

Section: Resultsmentioning

confidence: 99%

Inhibition of catechol-O-methyltransferase by natural pentacyclic triterpenes: structure–activity relationships and kinetic mechanism

Wang

Wei

Fan

et al. 2021

Journal of Enzyme Inhibition and Medicinal Chemistry

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Inhibitors of COMT are clinically used for the treatment of Parkinson's disease. Here, we report the first natural pentacyclic triterpenoid-type COMT inhibitors and their structure-activity relationships and inhibition mechanism. The most potent compounds were found to be oleanic acid, betulinic acid and celastrol with IC 50 values of 3.89-5.07 lM, that acted as mixed (uncompetitive plus non-competitive) inhibitors of COMT, representing a new skeleton of COMT inhibitor. Molecular docking suggested that they can specifically recognise and bind with the unique hydrophobic residues surrounding the catechol pocket. Furthermore, oleanic acid and betulinic acid proved to be less disruptive of mitochondrial membrane potential (MMP) compared to tolcapone, thus reducing the risk of liver toxicity. These findings could be used to produce an ideal lead compound and to guide synthetic efforts in generating related derivatives for further preclinical testing.

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Section: Resultsmentioning

confidence: 99%

Inhibition of catechol-O-methyltransferase by natural pentacyclic triterpenes: structure–activity relationships and kinetic mechanism

Wang

Wei

Fan

et al. 2021

Journal of Enzyme Inhibition and Medicinal Chemistry

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“…Several seminal reports have paved the ground for exploring the organization of mitochondria membranes and the co-localization of mitochondria proteins by super-resolution microscopy, at increased spatial sensitivity, partially bypassing the diffraction limits of visible light microscopy [28][29][30] and in live cells 30,31 . Thorough two-color 2D-STED analysis was applied to colocalize protein in the mitochondria membrane [32][33][34] , and the role of MIC26 and MIC27 in the stability and integration of the remaining MICOS subunits was studied by combining STED nanoscopy, and blue-native gel electrophoresis 35 .…”

Section: Discussionmentioning

confidence: 99%

Mitochondria respiratory complexes and supercomplexes co-exist simultaneously in mammalian cells

Enríquez

Martín‐García

Cabrera

et al. 2022

Preprint

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The mitochondrial electron transport chain was understood as four respiratory complexes (RCs) independently moving in the mitochondrial inner membrane and electrons transferred between them by CoQ and cytochrome c1,2. The disruption of the mitochondrial membranes with mild detergents followed by blue native electrophoresis (BNGE), allowed visualization of RCs as independent entities, but also associated forming a variety of supercomplexes (SCs)1,3. This led to the proposal that RCs are not moving free entities but instead they super-assembly into structures, called supercomplexes (SCs) or respirasomes3. The use of detergents raised concerns on the existence of either free RCs or SCs in the cellular context4, and does not allow determining the relevance of additional SCs. To address this controversy, it was proposed the plasticity model that considered that both RCs and SCs co-existed in a dynamic organization1,5,6. Over the last 20 years the reality of respiratory supercomplexes was confirmed by functional3,6–8, genetic6,9 and structural10–15 studies. However, the co-existence of free RCs and SCs is still unanswered. We applied multi-color Stimulated Emission Depletion (STED) microscopy in whole cells in a panel of cell lines: mtDNA depleted cells (p0), complex III (CYTbM) or complex IV (COX10KO) depleted cell lines and their isogenic controls. We demonstrate the co-existence of RCs and SCs in large cell areas. Our observations support the plasticity organization model of the mitochondrial respiratory chain were RCs and SCs co-exist and reveal an unexpected heterogeneous distribution of RCs and SCs within mitochondria in-situ.

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Shadow Electrochemiluminescence Microscopy of Single Mitochondria

Colin

Descamps

et al. 2021

Angewandte Chemie

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Mitochondria are the subcellular bioenergetic organelles. The analysis of their morphology and topology is essential to provide useful information on their activity and metabolism. Herein, we report a label‐free shadow electrochemiluminescence (ECL) microscopy based on the spatial confinement of the ECL‐emitting reactive layer to image single living mitochondria deposited on the electrode surface. The ECL mechanism of the freely‐diffusing [Ru(bpy)3]2+ dye with the sacrificial tri‐n‐propylamine coreactant restrains the light‐emitting region to a micrometric thickness allowing to visualize individual mitochondria with a remarkable sharp negative optical contrast. The imaging approach named “shadow ECL” (SECL) reflects the negative imprint of the local diffusional hindrance of the ECL reagents by each mitochondrion. The statistical analysis of the colocalization of the shadow ECL spots with the functional mitochondria revealed by classical fluorescent biomarkers, MitoTracker Deep Red and the endogenous intramitochondrial NADH, validates the reported methodology. The versatility and extreme sensitivity of the approach are further demonstrated by visualizing single mitochondria, which remain hardly detectable with the usual biomarkers. Finally, by alleviating problems of photobleaching and phototoxicity associated with conventional microscopy methods, SECL microscopy should find promising applications in the imaging of subcellular structures.

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A Multi‐responsive Fluorescent Probe Reveals Mitochondrial Nucleoprotein Dynamics with Reactive Oxygen Species Regulation through Super‐resolution Imaging

Cited by 7 publications

References 48 publications

Inhibition of catechol-O-methyltransferase by natural pentacyclic triterpenes: structure–activity relationships and kinetic mechanism

Inhibition of catechol-O-methyltransferase by natural pentacyclic triterpenes: structure–activity relationships and kinetic mechanism

Mitochondria respiratory complexes and supercomplexes co-exist simultaneously in mammalian cells

Shadow Electrochemiluminescence Microscopy of Single Mitochondria

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