Dataset Information

OTUD7B promotes cardiac hypertrophy by deubiquitinating SERCA2a

ABSTRACT: Cell samples were lysed in lysis buffer (8 M urea, 1% protease inhibitor cocktail). The supernatant was collected and the protein concentration was determined with BCA kit according to the manufacturer’s instructions. The peptides were obtained by digesting the protein with trypsin. Then, peptides were used for ubiquitin peptides enrichment by pre-washed antibody beads (PTM-1104, PTM Bio, Hangzhou, China). The subsequent quantitative ubiquitinome analysis was carried out by PTM Bio.

ORGANISM(S): Rattus Norvegicus

SUBMITTER: Guang Liang

PROVIDER: PXD068918 | iProX | Sat Sep 27 00:00:00 BST 2025

REPOSITORIES: iProX

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Publications

Cardiomyocyte-derived OTUD7B promotes cardiac hypertrophy by deubiquitinating SERCA2a.

Huang Zhuqi Z Han Xue X Hou Yuxing Y Wang Xingchen X Qiu Fuyu F Gong Yingchao Y Chattipakorn Nipon N Fu Guosheng G Liang Guang G Lai Dongwu D

Theranostics 20260204 8

<b>Rationale:</b> Pathological cardiac hypertrophy, triggered by persistent neurohumoral or hemodynamic stress, is a key precursor of ventricular dysfunction and heart failure. Deubiquitinating enzymes (DUBs) have emerged as critical regulators of cardiovascular biology. This study examined the function of a DUB, ovarian tumor domain-containing 7B (OTUD7B), in cardiac hypertrophy. <b>Methods:</b> Cardiomyocyte-specific OTUD7B knockout and overexpression mouse models were generated to evaluate my ...[more]

PMID: 41695487

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Project description:For proteomics analysis, testes of Ube2j2-/- and WT mice were collected on ice, washed once with pre-cooled PBS, dried with absorbent paper, and immediately placed in liquid nitrogen, dry ice, or at -80 ℃. Samples were packed in dry ice and transported to Hangzhou Jingjie Biotechnology Company (PTM Biolabs, Inc.) for whole protein 4D label-free quantitative proteomic determination. Testes of Ube2j2-/- and WT mice were subjected to LC-MS/MS analysis following a standard protocol. The sample was grinded and mixed with lysis buffer (8 M urea, 1% Protease Inhibitor Cocktail III (Merck Millipore, 539134) dissolved in aqueous solution containing buffer salts), followed by sonication on ice using a high intensity ultrasonic processor (Scientz). The remaining debris was removed by centrifugation at 12,000 g at 4 °C for 10 min. Then, the supernatant was collected and the protein concentration was determined with BCA kit (Beyotime Biotechnology, P0011) according to the manufacturer’s instructions. The sample was slowly added to a final concentration of 20% (m/v) trichloroacetic acid (Sigma-Aldrich, T4885) to precipitate proteins, then mixed by vortexing and incubated for 2 h at 4 °C. The precipitate was collected by centrifugation at 4500 x g for 5 min at 4°C. The precipitated protein was washed 3 times with pre-cooled acetone and dried for 1 min. Protein samples were then redissolved in 200 mM TEAB (Triethylammonium bicarbonate buffer, Sigma-Aldrich, T7408) and ultrasonically dispersed. Trypsin was added at 1:50 trypsin-to-protein mass ratio for the first digestion overnight. The sample was reduced with 5 mM dithiothreitol for 60 min at 37 °C and alkylated with 11 mM iodoacetamide for 45 min at room temperature in darkness. Finally, the peptides were desalted using a Strata X SPE column (Phenomenex). The tryptic peptides were fractionated into fractions by high pH reverse-phase HPLC using Agilent 300Extend C18 column (5 μm particles, 4.6 mm ID, 250 mm length). The released peptides were subjected to NSI source followed by tandem mass spectrometry (MS/MS) in Q ExactiveTM Plus (Thermo) coupled online to the UPLC. Peptides were then selected for MS/MS using NCE setting as 28 and the fragments were detected in the Orbitrap at a resolution of 17,500. Maxquant 1.6.15.0 software was used for spectra analysis, database searches, and peptide identification.

Project description:This batch of data is for three independent projects. Since in this three projects, the mouse strains are all CD1/ICR, the sex are both female, and the mouse ages are all two-month-old, three projects shared control data to save expenditure legitimately. But in our hand we couldn't segregate the raw data into two parts corresponding to two projects, so we uploads all raw data. However, we also provided researcher-analyzable excel files including the expression of all phosphorylated proteins. The concise description of these three projects are as follow: Project 1: We found that in Chronic Resistant Stress (CRS)-treated mice, the fertility and oocyte quality significantly decreased, and plenty of biochemical indexes significantly changed, so we guessed that the overall physical health of CRS mice was severely impacted. As one major strategy to investigate the mechanism how CRS impact the physical health, We chose TMT-labeled quantitative ovarian phosphoproteomics service from Hangzhou PTM Bio Co. We sent two repeats of two-month-old CRS CD1/ICR mouse ovaries and two repeats of control ovaries for characterization of differentially phosphorylated proteins. Each repeat contained about 60 ovaries from 30 female mice, about 300 mg. Project 2: We found that in High Salt (HS)-treated mice, the fertility and oocyte quality significantly decreased, and plenty of biochemical indexes significantly changed, so we guessed that the overall physical health of HS mice was severely impacted. As one major strategy to investigate the mechanism how HS impact the physical health, We chose TMT-labeled Quantitative ovarian phosphoproteomics service from Hangzhou PTM Bio Co. We sent two repeats of two-month-old HS CD1/ICR mouse ovaries and two repeats of control ovaries for characterization of differentially phosphorylated proteins. Each repeat contained about 60 ovaries from 30 female mice, about 300 mg. Project 3: We found that in arecoline (Are)-treated mice, the fertility and oocyte quality significantly decreased, and plenty of biochemical indexes significantly changed, so we guessed that the overall physical health of Are mice was severely impacted. As one major strategy to investigate the mechanism how Are impact the physical health, We chose TMT-labeled Quantitative ovarian phosphoproteomics service from Hangzhou PTM Bio Co. We sent two repeats of two-month-old Are CD1/ICR mouse ovaries and two repeats of control ovaries for characterization of differentially phosphorylated proteins. Each repeat contained about 60 ovaries from 30 female mice, about 300 mg. Two repeats of control samples and data were shared among project 1, 2 and 3. Gross protein levels were also examined and used to normalize the protein phosphorylation level.

Dataset Information

OTUD7B promotes cardiac hypertrophy by deubiquitinating SERCA2a

Publications

Cardiomyocyte-derived OTUD7B promotes cardiac hypertrophy by deubiquitinating SERCA2a.

Similar Datasets

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