Summary of Study ST004350
This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR002759. The data can be accessed directly via it's Project DOI: 10.21228/M81G1D This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.
| Study ID | ST004350 |
| Study Title | The metabolic effects of succinylation and desuccinylation of HADHB at lysine 292 |
| Study Summary | To elucidate the metabolic effects of succinylation and desuccinylation at lysine 292 (K292) of HADHB, we generated stable H9C2 rat cardiomyocyte cell lines expressing HADHB mutants. Specifically, lysine 292 was substituted with arginine (K292R) to mimic the desuccinylated state and with glutamic acid (K292E) to mimic the negatively charged succinylated modification. Non-targeted metabolomic profiling was performed on three groups—wild-type (WT), desuccinylation-mimic (K292R), and succinylation-mimic (K292E)—with six biological replicates per group. The metabolomic analysis revealed that succinylation at K292 of HADHB in cardiomyocytes leads to distinct metabolic alterations, particularly affecting the fatty acid β-oxidation pathway. |
| Institute | Nanchang University Second Affiliated Hospital |
| Last Name | Zeng |
| First Name | Zhimin |
| Address | nanchang, nanchang, jiangxi, 330006, China |
| 2zm@163.com | |
| Phone | 15979727903 |
| Submit Date | 2025-11-07 |
| Num Groups | 3 |
| Total Subjects | 6 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | mzML |
| Analysis Type Detail | LC-MS |
| Release Date | 2025-12-05 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR002759 |
| Project DOI: | doi: 10.21228/M81G1D |
| Project Title: | Untargeted metabolomic analysis of the metabolic effects of HADHB lysine 292 succinylation and desuccinylation |
| Project Summary: | HADHB succinylation status variations may influence metabolic states. We generated stable H9C2 rat cardiomyocyte cell lines expressing HADHB mutants. Untargeted metabolomics revealed that the succinylation and desuccinylation states at lysine 292 of HADHB may affect the Beta Oxidation of Very Long Chain Fatty Acids. |
| Institute: | Nanchang University Second Affiliated Hospital |
| Last Name: | Zeng |
| First Name: | Zhimin |
| Address: | nanchang, nanchang, jiangxi, 330006, China |
| Email: | 2zm@163.com |
| Phone: | 15979727903 |
Subject:
| Subject ID: | SU004509 |
| Subject Type: | Mammal |
| Subject Species: | Rattus norvegicus |
| Taxonomy ID: | 10116 |
| Genotype Strain: | HADHB-K292R、HADHB-K292E |
Factors:
Subject type: Mammal; Subject species: Rattus norvegicus (Factor headings shown in green)
| mb_sample_id | local_sample_id | Sample source | Genotype |
|---|---|---|---|
| SA517166 | E6 | H9C2 cells | HADHB-K292E |
| SA517167 | E5 | H9C2 cells | HADHB-K292E |
| SA517168 | E4 | H9C2 cells | HADHB-K292E |
| SA517169 | E3 | H9C2 cells | HADHB-K292E |
| SA517170 | E2 | H9C2 cells | HADHB-K292E |
| SA517171 | E1 | H9C2 cells | HADHB-K292E |
| SA517172 | R5 | H9C2 cells | HADHB-K292R |
| SA517173 | R6 | H9C2 cells | HADHB-K292R |
| SA517174 | R4 | H9C2 cells | HADHB-K292R |
| SA517175 | R3 | H9C2 cells | HADHB-K292R |
| SA517176 | R2 | H9C2 cells | HADHB-K292R |
| SA517177 | R1 | H9C2 cells | HADHB-K292R |
| SA517178 | C2 | H9C2 cells | Wild-type |
| SA517179 | C6 | H9C2 cells | Wild-type |
| SA517180 | C5 | H9C2 cells | Wild-type |
| SA517181 | C4 | H9C2 cells | Wild-type |
| SA517182 | C3 | H9C2 cells | Wild-type |
| SA517183 | C1 | H9C2 cells | Wild-type |
| Showing results 1 to 18 of 18 |
Collection:
| Collection ID: | CO004502 |
| Collection Summary: | H9C2 cells were purchased from Pronova (Punosai/Pronova-style naming), and a lentiviral system designed by Hanheng was used to introduce a lysine-to-arginine mutation at position 292 in HADHB (K292R) to generate a non-succinylated mutant, and a lysine-to-glutamate mutation at the same site (K292E) to mimic succinylation. Stable cell lines were established for HADHB-K292R (non-succinylated mutant) and HADHB-K292E (succiny-lation-mimetic). Seed six replicates per group from three growth conditions into 10 cm culture dishes. When the cells reach confluence, wash 2–3 times with pre-chilled PBS, add 1 mL of chromatography-grade methanol to each dish, detach the cells with a scraper, and transfer them to a 2 mL cryovial. Snap-freeze in liquid nitrogen for 15 minutes and then store at −80°C. Non-targeted metabolomics sequencing was performed on the three groups. |
| Sample Type: | Rat H9C2 cell line |
Treatment:
| Treatment ID: | TR004518 |
| Treatment Summary: | No treatment. |
Sample Preparation:
| Sampleprep ID: | SP004515 |
| Sampleprep Summary: | A 500 μL solution (Methanol: Water = 4:1, V/V) containing internal standard was added into the sample and vortexed for 3 min. The sample was placed in liquid nitrogen for 5 min and on the dry ice for 5 min, and then thawed on ice and vortexed for 2 min. This freeze-thaw circle was repeated three times in total. The sample was centrifuged at 12000 rpm for 10 min (4 °C). All of the supernatant was transferred and concentrated. A 100 μL solution (Methanol: Water = 7:3, V/V) was used to reconstitute the sample. Then the sample was vortexd for 3 minutes, and sonicate for 10 minutes in an ice bath. The sample was then centrifuged at 12000 rpm for 3 min (4 °C). A 80 μL aliquots of supernatant were transferred for LC-MS analysis. |
Chromatography:
| Chromatography ID: | CH005511 |
| Chromatography Summary: | All samples were for two LC/MS methods. One aliquot was analyzed using positive ion conditions and was eluted from T3 column (Waters ACQUITY Premier HSS T3 Column 1.8 µm, 2.1 mm * 100 mm) using 0.1 % formic acid in water as solvent A and 0.1 % formic acid in acetonitrile as solvent B in the following gradient: 5 to 20 % in 2 min, increased to 60 % in the following 3 mins, increased to 99 % in 1 min and held for 1.5 min, then come back to 5 % mobile phase B witnin 0.1 min, held for 2.4 min. The analytical conditions were as follows, column temperature, 40 °C; flow rate, 0.4 mL/min; injection volume, 4 μL; Another aliquot was using negative ion conditions and was the same as the elution gradient of positive mode. |
| Chromatography Comments: | Reversed-phase separation was performed on a Waters ACQUITY UPLC HSS T3 column (1.8 µm, 2.1 × 100 mm). The mobile phases consisted of (A) water with 0.1% formic acid and (B) acetonitrile. The flow rate was 0.3 mL/min. The gradient started at 2% B, increased linearly to 98% B over 15 min, held for 2 min, and then returned to initial conditions for re-equilibration. The column temperature was maintained at 40 °C, and the injection volume was 2 µL. |
| Instrument Name: | Thermo Vanquish |
| Column Name: | Waters ACQUITY UPLC HSS T3 (100 x 2.1mm,1.8um) |
| Column Temperature: | 40 |
| Flow Gradient: | 0–2 min: 5%–20% B; 2–5 min: 20%–60% B; 5–6 min: 60%–99% B; 6–7.5 min: 99% B; 7.5–7.6 min: 99%–5% B; 7.6–10 min: 5% B (re-equilibration) |
| Flow Rate: | 0.4 mL/min |
| Solvent A: | 100% water; 0.1% acetic acid |
| Solvent B: | 100% acetonitrile; 0.1% acetic acid |
| Chromatography Type: | Reversed phase |
Analysis:
| Analysis ID: | AN007261 |
| Analysis Type: | MS |
| Chromatography ID: | CH005511 |
| Num Factors: | 3 |
| Num Metabolites: | 1154 |
| Units: | Peak area |
| Analysis ID: | AN007262 |
| Analysis Type: | MS |
| Chromatography ID: | CH005511 |
| Num Factors: | 3 |
| Num Metabolites: | 779 |
| Units: | Peak Area |
