Direct tests of cytochrome c and c 1 functions in the electron transport chain of malaria parasites DOI Creative Commons
Tanya J. Espino-Sanchez, Henry Wienkers,

Rebecca G. Marvin

et al.

Proceedings of the National Academy of Sciences, Journal Year: 2023, Volume and Issue: 120(19)

Published: May 1, 2023

The mitochondrial electron transport chain (ETC) of Plasmodium malaria parasites is a major antimalarial drug target, but critical cytochrome (cyt) functions remain unstudied and enigmatic. Parasites express two distinct cyt c homologs (c c-2) with unusually sparse sequence identity uncertain fitness contributions. P. falciparum c-2 the most divergent eukaryotic homolog currently known has features predicted to be incompatible canonical ETC function. We tagged both related c1 for inducible knockdown. Translational repression was lethal parasites, which died from dysfunction impaired ubiquinone recycling. In contrast, knockdown or knockout had little impact on blood-stage growth, indicating that rely fully more conserved Biochemical structural studies revealed are hemylated by holocytochrome synthase, UV-vis absorbance EPR spectra strongly suggest an open active site in heme stably coordinated only single axial amino acid ligand can bind exogenous small molecules. These provide direct dissection identify highly molecular adaptations defy role evolution.

Language: Английский

Spatial-proteomics reveals phospho-signaling dynamics at subcellular resolution DOI Creative Commons
Ana Martínez‐Val, Dorte B. Bekker‐Jensen, Sophia Steigerwald

et al.

Nature Communications, Journal Year: 2021, Volume and Issue: 12(1)

Published: Dec. 7, 2021

Abstract Dynamic change in subcellular localization of signaling proteins is a general concept that eukaryotic cells evolved for eliciting coordinated response to stimuli. Mass spectrometry-based proteomics combination with fractionation can provide comprehensive maps spatio-temporal regulation protein networks cells, but involves laborious workflows does not cover the phospho-proteome level. Here we present high-throughput workflow based on sequential cell profile global proteome and dynamics across six distinct fractions. We benchmark by studying EGFR phospho-signaling vitro HeLa vivo mouse tissues. Finally, investigate stress signaling, revealing cellular relocation ribosomal hypertonicity muscle contraction. Proteomics data generated this study be explored through https://SpatialProteoDynamics.github.io .

Language: Английский

Citations

65
Emerging Chemical Biology of Protein Persulfidation DOI Creative Commons
Thibaut Vignane, Miloš R. Filipović

Antioxidants and Redox Signaling, Journal Year: 2023, Volume and Issue: 39(1-3), P. 19 - 39

Published: June 8, 2023

Significance: Protein persulfidation (the formation of RSSH), an evolutionarily conserved oxidative posttranslational modification in which thiol groups cysteine residues are converted into persulfides, has emerged as one the main mechanisms through hydrogen sulfide (H2S) conveys its signaling. Recent Advances: New methodological advances persulfide labeling started unraveling chemical biology this and role (patho)physiology. Some key metabolic enzymes regulated by persulfidation. RSSH levels important for cellular defense against injury, they decrease with aging, leaving proteins vulnerable to damage. Persulfidation is dysregulated many diseases. Critical Issues: A relatively new field signaling protein still unanswered questions: mechanism(s) transpersulfidation identification "protein persulfidases," improvement methods monitor changes identify targets, understanding controls (patho)physiological functions. Future Directions: Deep mechanistic studies using more selective sensitive techniques will provide high-resolution structural, functional, quantitative, spatiotemporal information on dynamics help better how H2S-derived affects structure function health disease. This knowledge could pave way targeted drug design a wide variety pathologies. Antioxid. Redox Signal. 39, 19-39.

Language: Английский

Citations

37
Modular Diazo Compound for the Bioreversible Late-Stage Modification of Proteins DOI
Joomyung V. Jun, Yana Petri,

Lucas W. Erickson

et al.

Journal of the American Chemical Society, Journal Year: 2023, Volume and Issue: 145(12), P. 6615 - 6621

Published: March 15, 2023

We introduce a versatile strategy for the bioreversible modification of proteins. Our is based on tricomponent molecule, synthesized in three steps, that incorporates diazo moiety chemoselective esterification carboxyl groups, pyridyl disulfide group late-stage functionalization with thiolated ligands, and self-immolative carbonate esterase-mediated cleavage. Using cytochrome c (Cyt c) green fluorescent protein (GFP) as models, we generated conjugates modified diverse domains cellular delivery include small targeting cell-penetrating peptides (CPPs), large polysaccharide. As proof concept, used our to effect proteins into cytosol live mammalian cells presence serum. The functional Cyt c, which induces apoptosis, highlighted advantage conjugation versus irreversible an amino group. ease utility this traceless provide new opportunities chemical biologists.

Language: Английский

Citations

34
The flexible chain: regulation of structure and activity of ETC complexes defines rate of ATP synthesis and sites of superoxide generation DOI

Zhanna V. Bochkova,

А. А. Байжуманов,

A. I. Yusipovich

et al.

Biophysical Reviews, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 25, 2025

Language: Английский

Citations

1
Probing Structure and Reaction Dynamics of Proteins Using Time-Resolved Resonance Raman Spectroscopy DOI
David Buhrke, Peter Hildebrandt

Chemical Reviews, Journal Year: 2019, Volume and Issue: 120(7), P. 3577 - 3630

Published: Dec. 9, 2019

The mechanistic understanding of protein functions requires insight into the structural and reaction dynamics. To elucidate these processes, a variety experimental approaches are employed. Among them, time-resolved (TR) resonance Raman (RR) is particularly versatile tool to probe processes proteins harboring cofactors with electronic transitions in visible range, such as retinal or heme proteins. TR RR spectroscopy offers advantage simultaneously providing molecular structure kinetic information. various spectroscopic methods can cover wide dynamic range down femtosecond time regime have been employed monitoring photoinduced cascades, ligand binding dissociation, electron transfer, enzymatic reactions, un- refolding. In this account, we review achievements nearly 50 years research field, which also illustrates how role life science has changed from beginning until now. We outline methodological developments point out current limitations potential perspectives.

Language: Английский

Citations

72
New moonlighting functions of mitochondrial cytochromecin the cytoplasm and nucleus DOI Open Access
Katiuska González‐Arzola, Alejandro Velázquez‐Cruz, Alejandra Guerra‐Castellano

et al.

FEBS Letters, Journal Year: 2019, Volume and Issue: 593(22), P. 3101 - 3119

Published: Oct. 30, 2019

Cytochrome c (C ) is a protein that functions as an electron carrier in the mitochondrial respiratory chain. However, C has moonlighting roles outside mitochondria driving transition of apoptotic cells from life to death. When living are damaged, escapes its natural environment and, once cytosol, it binds other proteins form complex named apoptosome—a platform triggers caspase activation and further leads controlled cell dismantlement. Early released also inositol 1,4,5‐triphosphate receptors on ER membrane, which stimulates massive release mitochondria. Besides well‐characterized binding contributing proapoptotic , many novel targets have been recently described. Among them, histone chaperones were identified key partners following DNA breaks, indicating might modulate chromatin dynamics through competitive chaperones. In this article, we review ample set discovered antiapoptotic proteins—involved damage, transcription, energetic metabolism—reported interact with cytoplasm even nucleus upon breaks.

Language: Английский

Citations

66
Immobilization of l-asparaginase on gold nanoparticles for novel drug delivery approach as anti-cancer agent against human breast carcinoma cells DOI Creative Commons

Ali G. Al-Dulimi,

Ali Z. Al‐Saffar, Ghassan M. Sulaiman

et al.

Journal of Materials Research and Technology, Journal Year: 2020, Volume and Issue: 9(6), P. 15394 - 15411

Published: Oct. 23, 2020

l-asparaginase is an amidohydrolase enzyme that widely identified as one of the most potential anti cancerous drugs. Nevertheless, this drug poorly bioavailable and hence its pharmaceutical uses are limited. To improve bioactivity, was loaded on gold nanoparticles (GNPs) along with Arg-Gly-Asp (RGD) peptide direct to targeted cancer cells aim enhancing anticancer efficiency. Successful preparation GNPs conjugate (GNPs-PEG-l-asparaginase-RGD) verified delineated by employing UV–VIS spectrophotometer, FTIR, XRD, FE-SEM, TEM. Fourier Transform Infrared (FT-IR), X-Ray Diffraction (XRD), Field Emission Scanning Electron Microscopic (FE-SEM), Transmission Microscopy (TEM). The efficiency target distribute in MCF-7 evaluated using high fluorescent signals confirmed fluorescence microscopy. A variety parameters were tested investigate each compound toward vitro. demonstrated significant antioxidant effects tumor targeting efficacy distribution cells. It caused a decrease cell proliferation rate clonogenicity cells, while initiating apoptosis promoting cycle arrest at G2/M, flow cytometry analysis. These coupled upregulating pro-apoptotic p53 downregulating anti-apoptotic Bcl-2, which resulted alleviation mitochondrial membrane (MMP) and, thereby, secretion cytochrome c. outcomes present study propose feasibility for further development novel agent against

Language: Английский

Citations

63
SERS uncovers the link between conformation of cytochrome c heme and mitochondrial membrane potential DOI Open Access
Nadezda A. Brazhe, Evelina I. Nikelshparg,

А. А. Байжуманов

et al.

Free Radical Biology and Medicine, Journal Year: 2023, Volume and Issue: 196, P. 133 - 144

Published: Jan. 14, 2023

Language: Английский

Citations

21
MYG1 drives glycolysis and colorectal cancer development through nuclear-mitochondrial collaboration DOI Creative Commons
Jian‐Xiong Chen,

Shiyu Duan,

Yulu Wang

et al.

Nature Communications, Journal Year: 2024, Volume and Issue: 15(1)

Published: June 11, 2024

Metabolic remodeling is a strategy for tumor survival under stress. However, the molecular mechanisms during metabolic of colorectal cancer (CRC) remain unclear. Melanocyte proliferating gene 1 (MYG1) 3'-5' RNA exonuclease and plays key role in mitochondrial functions. Here, we uncover that MYG1 expression upregulated CRC progression highly expressed promotes glycolysis independent its activity. Mechanistically, nuclear recruits HSP90/GSK3β complex to promote PKM2 phosphorylation, increasing stability. transcriptionally activates MYC MYC-medicated glycolysis. Conversely, c-Myc also upregulates MYG1, driving CRC. Meanwhile, on one hand inhibits oxidative phosphorylation (OXPHOS), other blocks release Cyt c from mitochondria cell apoptosis. Clinically, patients with KRAS mutation show high indicating level poor prognosis. Targeting may disturb balance serve as potential target diagnosis treatment

Language: Английский

Citations

8
Real-Time Observation of Conformational Changes and Translocation of Endogenous Cytochrome c within Intact Mitochondria DOI

Jianhua Zhan,

Danyun Zeng,

Xiong Xiao

et al.

Journal of the American Chemical Society, Journal Year: 2024, Volume and Issue: 146(7), P. 4455 - 4466

Published: Feb. 9, 2024

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

Language: Английский

Citations

7