Browse
Submit Data
Databases
API
Help

Dataset Information

0 Views

0 Connections

0 Citations

0 Reanalyses

0 Downloads

Omics score: 0

Site-specific Identification and Quantification of Protein Glutathionylation under Ischemic Stress

ABSTRACT: Ischemia or ischemic reperfusion is well-known for its contribution to heart diseases. During ischemia and reperfusion, a flux of nutrients and oxygen to cardiomyocytes is altered, which causes a burst of ROS from mitochondria and other enzymes. Elevated levels of ROS can cause oxidative modifications of cardiac proteins, such as protein glutathionylation. Despite previous extensive studies, proteomic identification of glutathionylated proteins in cardiomyocytes under altered levels of nutrients and oxygen has been relatively limited. We have applied our clickable glutathione approach to HL-1 cardiomyocyte cell line under metabolic alterations of glucose, oxygen, and fatty acids to detect and identify proteins undergoing glutathionylation.

INSTRUMENT(S): Orbitrap Fusion

ORGANISM(S): Mus Musculus (mouse)

SUBMITTER: Maheeshi Yapa Abeywardana

LAB HEAD: Young-Hoon Ahn

PROVIDER: PXD024766 | Pride | 2021-08-13

REPOSITORIES: Pride

ACCESS DATA

Json Xml

Dataset's files

Source:

			Action	DRS
	2258_4-126A.csv	Csv
	2258_4-126A.raw	Raw
	2258_4-126B-a.csv	Csv
	2258_4-126B-a.raw	Raw
	2258_4-126B-b.csv	Csv

Items per page:

1 - 5 of 13

Publications

Identification and Quantification of Glutathionylated Cysteines under Ischemic Stress.

Yapa Abeywardana Maheeshi M Samarasinghe Kusal T G KTG Munkanatta Godage Dhanushka D Ahn Young-Hoon YH

Journal of proteome research 20210812 9

Ischemia reperfusion injury contributes to adverse cardiovascular diseases in part by producing a burst of reactive oxygen species that induce oxidations of many muscular proteins. Glutathionylation is one of the major protein cysteine oxidations that often serve as molecular mechanisms behind the pathophysiology associated with ischemic stress. Despite the biological significance of glutathionylation in ischemia reperfusion, identification of specific glutathionylated cysteines under ischemic s ...[more]

PMID: 34382403

Similar Datasets

Clickable glutathione-based Isotope Coded Affinity Label to quantify glutathionylation

Project description:Identification of cysteines with high oxidation susceptibility is important for understanding redox-mediated biological processes associated with health and disease. We developed a chemical proteomic strategy that helps find cysteines with high susceptibility to S-glutathionylation. Our strategy is based on the isotopically labelled clickable glutathione derivatives that can quantify relative levels of glutathionylated and reduced forms (SSG vs. SH) of cysteines in mass spectrometry, which is named 'Clickable Glutathione-based Isotope-coded Affinity-Tag (G-ICAT).' The G-ICAT approach was applied to the mouse C2C12 cell line upon addition of hydrogen peroxide, finding 1,518 glutathionylated cysteines and their relative levels of SSG over SH upon adding hydron peroxide. This chemical proteomic strategy will significantly contribute to identifying functional cysteines regulated by glutathionylation in redox signaling.

2024-03-07 | PXD034976 | Pride

McDougal2017 - Metabolism in ischemic cardiomyocytes

Project description:Heart disease remains the leading cause of death globally. Although reperfusion following myocardial ischemia can prevent death by restoring nutrient flow, ischemia/reperfusion injury can cause significant heart damage. The mechanisms that drive ischemia/reperfusion injury are not well understood; currently, few methods can predict the state of the cardiac muscle cell and its metabolic conditions during ischemia. Here, we explored the energetic sustainability of cardiomyocytes, using a model for cellular metabolism to predict the levels of ATP following hypoxia. We modeled glycolytic metabolism with a system of coupled ordinary differential equations describing the individual metabolic reactions within the cardiomyocyte over time. Reduced oxygen levels and ATP consumption rates were simulated to characterize metabolite responses to ischemia. By tracking biochemical species within the cell, our model enables prediction of the cell’s condition up to the moment of reperfusion. The simulations revealed a distinct transition between energetically sustainable and unsustainable ATP concentrations for various energetic demands. Our model illustrates how even low oxygen concentrations allow the cell to perform essential functions. We found that the oxygen level required for a sustainable level of ATP increases roughly linearly with the ATP consumption rate. An extracellular O2 concentration of ~0.007 mM could supply basic energy needs in non-beating cardiomyocytes, suggesting that increased collateral circulation may provide an important source of oxygen to sustain the cardiomyocyte during extended ischemia. Our model provides a time-dependent framework for studying various intervention strategies to change the outcome of reperfusion.

2024-09-02 | BIOMD0000000961 | BioModels

In-vitro glutathionylation of SMYD2

Project description:Glutathionylation is an important post-translational modification and developing tools to identify which cysteine residues on proteins are glutathionylated in response to different physiological conditions is necessary. This project utilizes an approach consisting of azido-glutathione, a single purified protein, and an oxidant to form disulfide bonds between the azide modified glutathione and the cysteine residues of the protein. After this the reaction is quenched with iodoacetamide, subjected to a click reaction with a chemically cleavable alkyne conjugated biotin, and digested with trypsin. These peptide fragments are eluted from streptavidin beads and subjected to LC-MS/MS to determine which cysteine residues are glutathionylated in-vitro.

2019-04-03 | PXD011123 | Pride

Lung transplantation-induced cold ischemia–reperfusion injury in the murine lung

Project description:Cold ischemia-reperfusion induced injury contributes to poor lung transplant outcomes. We used transcriptome sequencing to study the biological response of mouse lungs to the cold ischemia-reperfusion process. Mouse orthotopic left LTx was performed with standard cuff techniques. Briefly, the donor lungs were recovered after being flushed with 10ml low potassium dextran (LPD) solution and inflated with 50% oxygen. Cold ischemia was induced by storing donor lungs in 20ml LPD at 4°C for 24 hours. Then, the left donor lung was cuffed and implanted into recipients within 45 minutes. After the 4-hour reperfusion, the recipient mice were sacrificed and the transplanted lungs were collected.

2023-01-20 | GSE222957 | GEO

Integrated stress response couples mitochondria protein translation with oxidative stress control

Project description:Under stress conditions, cells elicit integrated stress response (ISR) to cope with intracellular and extracellular disturbances. However, its role in ischemic heart disease remains to be elucidated. Here, we show that oxygen deprivation in cardiomyocytes triggers significant changes in protein translation. Importantly, ischemia and ischemia/reoxygenation leads to suppression of protein synthesis, which is caused by activation of the PERK/eIF2α axis of ISR. At the functional level, cardiac specific elimination of PERK exacerbates cardiac response to ischemia/reperfusion whereas selective activation of PERK in the heart confers cardioprotection against reperfusion injury. Mechanistically, PERK-mediated improvement in cardiomyocyte survival depends on suppression of protein synthesis and consequently relieves energetic demand on mitochondria. We went further to show that mitochondrial complex components are targeted by protein translation suppression, which significantly diminishes mitochondria-associated production of reactive oxygen species. Indeed, pharmacological activation of ISR protects the heart from ischemia/reperfusion damage, even after the release of occluded coronary artery, highlighting clinical significance for myocardial infarction. Taken together, these findings suggest that ISR improves cell survival through selectively suppressing mitochondrial protein synthesis and reducing oxidative stress in ischemic heart disease.

2022-10-01 | GSE177078 | GEO

Ischemic Pre- and Post Conditioning has pronounced effects on Gene Expression Profiles in the Rat Liver after Ischemia/Reperfusion

Project description:Ischemia/reperfusion injuries is a known complication to hepatic surgery. Ischemic pre- (IPC) and postconditioning (IPO) protects the liver against ischemia/reperfusion-injuries. Expression profiling were performed on liver biopsies seeking to identify molecular mediators of the protective properties. 48 rats were divided into 5 groups; sham (n=8), IRI (n=10), IPC (n=10), IPO (n=10) and IPC+IPO (n=10). All rats except sham rats were subjected to 30 min of total liver ischemia and 30 min of reperfusion before liver biopsies were sampled. In the IPC group, liver ischemia was preceded by 10 min of hepatic ischemia, followed by 10 min of reperfusion. IPO were performed by three cycles of 30 sec of reperfusion and 30 sec of ischemia, applied immediately after the 30 min of total liver ischemia. In the IPC+IPO group the two interventions were combined.

2011-09-30 | E-GEOD-24430 | biostudies-arrayexpress

Transcription profiling of rat NRVMs response to ischemia and reperfusion events using Coverslip Hypoxia

Project description:NRVMs were subjected to varying durations of ischemia or ischemia+reperfusion using coverslip hypoxia. Expression profiling was used to identify genes that are differentially regulated in either event. We used microarrays to detail the global program of gene expression underlying ischemia and reperfusion using Coverslip Hypoxia and identified distinct classes of genes regulated during these processes. Experiment Overall Design: RNA was extracted from NRVMs subjected to varying durations of ischemia or ischemia+reperfusion and hybridized to rat genome Affymetrix arrays.

2008-06-15 | E-GEOD-6701 | biostudies-arrayexpress

Global microRNA expression profiling of mouse livers following ischemia-reperfusion injury at different stages. [miRNA]

Project description:Hepatic ischemia reperfusion injury is a dynamic process consisting of two stages: ischemia and reperfusion, and triggers a cascade of physiological and biochemical events. Given the important role of microRNAs in regulating gene expression, we analyzed gene expression changes in mouse livers at sham control, ischemia stage, and reperfusion stage. We generated global expression profiles of microRNA and mRNA genes in mouse livers subjected to ischemia reperfusion injury at the three stages, respectively. Comparison analysis showed that reperfusion injury had a distinct expression profile whereas the ischemia sample and the sham control were clustered together. Consistently, there are 69 differentially expressed microRNAs between the reperfusion sample and the sham control whereas 28 differentially expressed microRNAs between the ischemia sample and the sham control. We further identified two modes of microRNA expression changes in ischemia reperfusion injury. Functional analysis of both the differentially expressed microRNAs in the two modes and their target mRNAs revealed that ischemia injury impaired mitochondria function, nutrient consumption, and metabolism process. In contrast, reperfusion injury led to severe tissue inflammation that is predominantly an innate-immune response in the ischemia reperfusion process. Our staged analysis of gene expression profiles provides new insights into regulatory mechanisms of microRNAs in mouse hepatic ischemia reperfusion injury.

2015-10-01 | GSE72314 | GEO

Global microRNA expression profiling of mouse livers following ischemia-reperfusion injury at different stages. [mRNA]

2015-10-01 | GSE72312 | GEO

Differential expression of mRNAs in the heart of ischemia-reperfusion mice

Project description:To identify the role of mRNA during myocardial ischemia-reperfusion in mice, we have employed high-throughput sequencing to detect mRNA expression. Samples were collected from the control group and the ischemia reperfusion groups , with 5 samples per group. The candidate mRNA that may affect the process of myocardial ischemia-reperfusion was screened by comparing the ischemia-reperfusion group and the control group.

2023-11-15 | GSE223343 | GEO