SERS uncovers the link between conformation of cytochrome c heme and mitochondrial membrane potential

Brazhe, Nadezda A.; Nikelshparg, Evelina I.; Baizhumanov, A. A.; Grivennikova, Vera G.; Semenova, Anna A.; Novikov, Sergey M.; Volkov, Valentyn S.; Arsenin, Aleksey V.; Yakubovsky, Dmitry I.; Evlyukhin, Andrey B.; Bochkova, Zhanna V.; Goodilin, Eugene A.; Максимов, Г. В.; Sosnovtseva, Olga; Rubin, Andrey B.

doi:10.1016/j.freeradbiomed.2023.01.013

Cited by 16 publications

(11 citation statements)

References 47 publications

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“…CCCP is a potent uncoupler of mitochondrial OXPHOS. It disrupts ATP synthesis by transporting protons across the mitochondrial inner membrane, interfering with the proton gradient. – The use of CCCP (0.7 μM, 15 min) causes a decrease in the mitochondrial membrane potential, which is observed from a reduced band intensity ratio of 750/1450 cm –1 in comparison to that of the control. Taking into account the obtained results, it can be postulated that the tested Rp forms covalent bonds with mitochondrial proteins, which slightly changes the mitochondrial activity [decrease in the ratio of 750/1450 cm –1 , as in the case of CCCP (Figure A,C)].…”

Section: Resultsmentioning

confidence: 99%

New Highly Sensitive and Specific Raman Probe for Live Cell Imaging of Mitochondrial Function

Pieczara,

Arellano Reyes,

Keyes

et al. 2024

ACS Sens.

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For Raman hyperspectral detection and imaging in live cells, it is very desirable to create novel probes with strong and unique Raman vibrations in the biological silent region (1800–2800 cm–1). The use of molecular probes in Raman imaging is a relatively new technique in subcellular research; however, it is developing very rapidly. Compared with the label-free method, it allows for a more sensitive and selective visualization of organelles within a single cell. Biological systems are incredibly complex and heterogeneous. Directly visualizing biological structures and activities at the cellular and subcellular levels remains by far one of the most intuitive and powerful ways to study biological problems. Each organelle plays a specific and essential role in cellular processes, but importantly for cells to survive, mitochondrial function must be reliable. Motivated by earlier attempts and successes of biorthogonal chemical imaging, we develop a tool supporting Raman imaging of cells to track biochemical changes associated with mitochondrial function at the cellular level in an in vitro model. In this work, we present a newly synthesized highly sensitive RAR-BR Raman probe for the selective imaging of mitochondria in live endothelial cells.

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Section: Resultsmentioning

confidence: 99%

New Highly Sensitive and Specific Raman Probe for Live Cell Imaging of Mitochondrial Function

Pieczara,

Arellano Reyes,

Keyes

et al. 2024

ACS Sens.

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“…Oxygen consumption rate (OCR) is one of the main indexes of respiration. It can be used to evaluate the metabolic status of mitochondria. , OCR of mitochondria was measured at Shanghai Fuda Testing Technology Group Co., Ltd. (Shanghai, China) by using a Seahorse Energy Metabolism Analyzer (Agilent Technologies, XFe24, USA). The mitochondria stock with a total protein concentration of 2 mg/mL was kept at 30 °C.…”

Section: Methodsmentioning

confidence: 99%

Real-Time Observation of Conformational Changes and Translocation of Endogenous Cytochrome c within Intact Mitochondria

Zhan,

Zeng,

Xiao

et al. 2024

J. Am. Chem. Soc.

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Cytochrome c (cyt c) is a multifunctional protein with varying conformations. However, the conformation of cyt c in its native environment, mitochondria, is still unclear. Here, we applied NMR spectroscopy to investigate the conformation and location of endogenous cyt c within intact mitochondria at natural isotopic abundance, mainly using widespread methyl groups as probes. By monitoring time-dependent chemical shift perturbations, we observed that most cyt c is located in the inner mitochondrial membrane and partially unfolded, which is distinct from its native conformation in solution. When suffering oxidative stress, cyt c underwent oxidative modifications due to increasing reactive oxygen species (ROS), weakening electrostatic interactions with the membrane, and gradually translocating into the inner membrane spaces of mitochondria. Meanwhile, the lethality of oxidatively modified cyt c to cells was reduced compared with normal cyt c. Our findings significantly improve the understanding of the molecular mechanisms underlying the regulation of ROS by cyt c in mitochondria. Moreover, it highlights the potential of NMR to monitor high-concentration molecules at a natural isotopic abundance within intact cells or organelles.

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“…As shown in Table 4, a total of three proteins were enriched to the oxidative phosphorylation pathway in the treated and control groups, in which mitochondrial cytochrome C protein expression was reduced, vacuolar ATP synthethase subunit e and ATP synthase subunit d protein expression were unchanged. Cytochrome C is an essential part of the mitochondrial respiratory chain, which is generated from two inactive precursor molecules, procytochrome C, and heme [33]. Since cytochrome C has a ferrous heme group, it can transfer electrons between respiratory chain complex enzyme III (cytochrome reductase) and respiratory chain complex enzyme IV (cytochrome oxidase) [34,35].…”

Section: Kegg Classification Analysismentioning

confidence: 99%

Discovery of a Novel Class of Acylthiourea-Containing Isoxazoline Insecticides against Plutella xylostella

Jiang

Luo

et al. 2023

Molecules

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Isoxazoline structures are widely found in natural products and are rich in biological activities. This study discloses the development of a series of novel isoxazoline derivatives by introducing acylthiourea fragments to access insecticidal activity. All synthetic compounds were examined for their insecticidal activity against Plutella xylostella, with results showing moderate to strong activity. Based on this, the structure–activity relationship analysis was carried out via the constructed three-dimensional quantitative structure–activity relationship model to further guide the structure optimization, resulting in the optimal compound 32. The LC50 of compound 32 against Plutella xylostella was 0.26 mg/L, demonstrating better activity than the positive control, ethiprole (LC50 = 3.81 mg/L), avermectin (LC50 = 12.32 mg/L), and compounds 1–31. The insect GABA enzyme-linked immunosorbent assay demonstrated that compound 32 might act on the insect GABA receptor, and the molecular docking assay further illustrated the mode of action of compound 32 with the GABA receptor. In addition, the proteomics analysis indicated that the action of compound 32 on Plutella xylostella was multi-pathway.

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SERS uncovers the link between conformation of cytochrome c heme and mitochondrial membrane potential

Cited by 16 publications

References 47 publications

New Highly Sensitive and Specific Raman Probe for Live Cell Imaging of Mitochondrial Function

New Highly Sensitive and Specific Raman Probe for Live Cell Imaging of Mitochondrial Function

Real-Time Observation of Conformational Changes and Translocation of Endogenous Cytochrome c within Intact Mitochondria

Discovery of a Novel Class of Acylthiourea-Containing Isoxazoline Insecticides against Plutella xylostella

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