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Quantitative proteomic analysis was developed on the colon of IF1-KO and control mice

ABSTRACT: We performed a quantitative proteomic analysis of colon tissue extracts from CRL and IF1-KO mice with isobaric tags for relative and absolute quantitation (iTRAQ) labeling method coupled to tandem mass spectrometry (MS/MS).

INSTRUMENT(S): LTQ Orbitrap Velos

ORGANISM(S): Mus Musculus (mouse)

TISSUE(S): Colon

SUBMITTER: Sonia Domínguez

LAB HEAD: José Manuel Cuezva

PROVIDER: PXD038107 | Pride | 2023-10-24

REPOSITORIES: Pride

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Dataset's files

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			Action	DRS
	020718_Fulvio_2_iTRAQ_F1.mgf	Mgf
	020718_Fulvio_2_iTRAQ_F1.raw	Raw
	020718_Fulvio_2_iTRAQ_F2.mgf	Mgf
	020718_Fulvio_2_iTRAQ_F2.raw	Raw
	020718_Fulvio_2_iTRAQ_F3.mgf	Mgf

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Publications

IF1 ablation prevents ATP synthase oligomerization, enhances mitochondrial ATP turnover and promotes an adenosine-mediated pro-inflammatory phenotype.

Domínguez-Zorita Sonia S Romero-Carramiñana Inés I Santacatterina Fulvio F Esparza-Moltó Pau B PB Simó Carolina C Del-Arco Araceli A Núñez de Arenas Cristina C Saiz Jorge J Barbas Coral C Cuezva José M JM

Cell death & disease 20230712 7

ATPase Inhibitory Factor 1 (IF1) regulates the activity of mitochondrial ATP synthase. The expression of IF1 in differentiated human and mouse cells is highly variable. In intestinal cells, the overexpression of IF1 protects against colon inflammation. Herein, we have developed a conditional IF1-knockout mouse model in intestinal epithelium to investigate the role of IF1 in mitochondrial function and tissue homeostasis. The results show that IF1-ablated mice have increased ATP synthase/hydrolase ...[more]

PMID: 37433784

Similar Datasets

Quantitative proteomic analysis of colon tissue extracts from CRL and IF1-KO mice

Project description:Quantitative proteomic analysis of colon tissue extracts from CRL and IF1-KO mice was performed with isobaric tags for relative and absolute quantitation (iTRAQ) labeling method coupled to tandem mass spectrometry (MS/MS).

2023-10-24 | PXD038151 | Pride

Genetic regulation of the ATP synthase in neurons modulates cognition

Project description:The mitochondrial ATP synthase produces ATP by oxidative phosphorylation and integrates different signals to regulate cellular functions and fate. The ATPase inhibitory factor 1 (IF1) is a structurally-disordered protein that inhibits the ATP synthase, contributing to metabolic reprogramming and signalling through mitochondrial reactive oxygen species (mtROS). mtROS regulate kinases and transcription factors in mitohormetic responses that favour adaptation to toxic insults2. IF1 is tissue-specifically expressed and in human and mouse brain is in molar excess over the ATP synthase. Herein, we have used genetic approaches to ablate or overexpress IF1 in neurons to investigate its role in brain functions. IF1 inhibits a fraction of the ATP synthase under physiological conditions and regulates respiration, mtROS production and mitochondrial structure. Transcriptomic, proteomic and metabolomic analyses indicate that IF1 regulates synaptic transmission and cognition. Ablation of IF1 impairs short-term memory whereas IF1 overexpression increases basal synaptic transmission and learning by mtROS-dependent activation of the extracellular signal-regulated kinases 1/2 (ERK 1/2). Overall, we show that IF1 dose plays a fundamental role in the regulation of neuronal function by controlling the fraction of inhibited ATP synthase that acts as source of mitohormetic mtROS, further emphasizing the ATP synthase/IF1 as promising targets to treat cognitive disorders.

2021-05-24 | PXD020262 | Pride

Skeletal muscle wt and OXPHOS inhibited 18 months-old mice

Project description:TMT sixplex Isobaric Mass Tagging (TAM) analysis was carried out in the CBMSO Protein Chemistry Facility (ProteoRed, PRB3-ISCIII and UAM University, Spain. Chronic skeletal muscle mitochondria dyshomeostasis drives tubular aggregate formation.

2022-08-12 | PXD026771 | Pride

Skeletal muscle wt and OXPHOS inhibited 18 months-old mice treated with edaravone

Project description:Chronic skeletal muscle mitochondria dyshomeostasis drives tubular aggregate formation. TMT sixplex Isobaric Mass Tagging analysis was carried out in the CBMSO Protein Chemistry Facility (ProteoRed, PRB3-ISCIII and UAM University, Spain.

2022-08-12 | PXD026722 | Pride

C2C12 myoblast WT vs ETFDH-ko TMT

Project description:ETFDH (electron transfer flavoprotein ubiquinone oxidoreductase) is a 64 kDa protein monomer located in the inner mitochondrial membrane, in charge of transferring the electrons received from the electron transfer flavoprotein ETF to the Coenzyme Q (Q). Pathological mutations in ETFDH lead to Multiple Acyl-CoA Dehydrogenase Deficiency (MADD; OMIM #231680). C2C12 cells lacking ETFDH were analysed by TMT analysis and compared to wt cells.

2023-10-24 | PXD041825 | Pride

Proteomic analysis of acute pancreatitis in TRP14-KO miceProteomic analysis of acute pancreatitis in TRP14-KO mice

Project description:We have studied pancreas samples in pancreas from a mouse model of acute pancreatitis, comparing wild-type animals and TRP14 KO animals. Samples are extracted with N-ethylmaleimide to block reduced cysteines, and no reducing agent is used, with a search including cysteinylated peptides. Acute pancreatitis was induced in 12 weeks-old mice (wild-type and KO for TRP14) by seven intraperitoneal injections of cerulein (50 µg/kg body weight) at 1 h intervals. Then, 1 h after the last injection, animals were euthanized under anesthesia with 3% isoflurane, exsanguinated, and the pancreas was immediately removed and processed.

2024-05-15 | PXD050765 | Pride

Proteins present in tears from control, MCI and Alzheimer disease patients

Project description:we investigated tear fluid as a novel source of disease-specific protein biomarkers for Alzheimer disease (AD) development using samples from patients suffering from mild cognitive impairment (MCI) and AD. Tear protein content was evaluated via liquid chromatography–mass spectrometry.

2019-11-04 | PXD014475 | Pride

Antimycin A-treated vs ETFDH-ko myoblasts TMT

Project description:The electron transfer flavoprotein dehydrogenase (ETFDH) is the bottleneck enzyme of fatty acid oxidation (FAO). We performed a TMT 6plex analysis on Antimycin A treated and ETFDH-ko myoblasts (compared to CRL).

2023-10-24 | PXD045588 | Pride

Proteome of the EPS of Lotmaria passim promastigotes

Project description:Honeybee health is a major ecological, agricultural and societal concern due to the critical role of these insects for plant reproduction and the massive losses of colonies observed within the last decade. The search for abiotic (e.g. pesticides) and biotic (e.g. pathogens) stressors is essential for understanding bee declines and design protection plans. Flagellated Trypanosomatid protozoan parasites and particularly Lotmaria passim, are widely distributed in honeybee colonies and have been associated with colony losses. However, little is known about their life cycles, routes for transmission and the strategies for survival inside and outside their hosts. Here we describe the proteome of L. passim Extracellular polymeric substances linked to secretion of Extracellular vesicles using liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). The EPS-EVs composition will provide clues on how these pathogens survive and spread in apiaries worldwide.

2024-02-15 | PXD040355 | Pride

Comparison of three methods for isolating human plasma EVs by LC-MSMS

Project description:The lack of standardized protocols for isolating extracellular vesicles (EVs), particularly from biobank-stored plasma, poses a substantial limitation for the study of biomarkers. Combining current isolation methods could enhance the specificity and purity of EVs isolation for their further use in diagnosis and personalized medicine. In this study, plasma samples from healthy individuals were subjected to extracellular vesicle isolation using ultracentrifugation (UC), size exclusion chromatography (SEC), and a combined approach of SEC and UC (SEC+UC). Characterization of the isolated EVs with NTA and TEM indicated that all isolation methods successfully isolated EVs, being not altered due to the long-term storage. Also, the western blot analysis confirmed the presence of exosome markers in all isolates but showed a higher expression of albumin in EVs-UC, suggesting potential plasma protein contamination. Proteomic analysis of samples from different isolation methods identified 542 proteins, with SEC+UC yielding the most complex proteome at 364 proteins. GO analysis of cellular component revealed differences in terms related to EVs and plasma, where SEC+UC proteins had the highest proportion of exosomal proteins. As a result of further study of the unique proteins detected in each isolation method, it was revealed that the SEC+UC method detected 181 unique proteins, demonstrating its superiority in the detection of low plasma concentration proteins. These findings support the robustness of the SEC+UC approach for EV isolation and proteomic studies due to its high efficiency and purity in isolating EVs. This study emphasizes the efficacy of the SEC+UC approach in obtaining highly pure and diverse EV populations suitable for comprehensive proteomic analysis with application to the discovery of biomarkers from biobank-stored plasma.

2024-07-03 | PXD049342 | Pride

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