Summary of Study ST003813
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 PR002385. The data can be accessed directly via it's Project DOI: 10.21228/M8BR77 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 | ST003813 |
| Study Title | Sulfur Amino Acid Restriction Enhances Exercise Capacity in Mice by Boosting Fat Oxidation in Muscle - extensor digitorum longus (EDL) versus soleus (Sol) on Con and SAAR diet metabolomics |
| Study Summary | Dietary restriction of the sulfur-containing amino acids methionine and cysteine (SAAR) improves body composition, enhances insulin sensitivity, and extends lifespan; benefits seen also with endurance exercise. Yet, the impact of SAAR on skeletal muscle remains largely unexplored. Here we demonstrate that one week of SAAR in sedentary, young, male mice increases endurance exercise capacity. Indirect calorimetry showed that SAAR increased lipid oxidation at rest and delayed the onset of carbohydrate utilization during exercise. Transcriptomic analysis revealed increased expression of genes involved in fatty acid catabolism especially in glycolytic muscle following SAAR. These findings were functionally supported by increased fatty acid circulatory turnover flux and muscle β-oxidation. Reducing lipid uptake from circulation through endothelial cell (EC)-specific CD36 deletion attenuated the running phenotype. Mechanistically, VEGF-signaling inhibition prevented exercise increases following SAAR, without affecting angiogenesis, implicating noncanonical VEGF signaling and EC CD36-dependent fatty acid transport in regulating exercise capacity by influencing muscle substrate availability. Here we performed metabolomics using EDL and Sol to evaluate the systemic effects of the diet on muscle metabolites. One week of dietary treatment was not sufficient to significantly alter a wide range of metabolites. However, known metabolites downstream of the methionine cycle, such as taurine, were significantly affected. Additionally, ophthalmic acid—which has been reported to compensate for glutathione loss under protein restriction (MacArthur et al., 2022; https://doi.org/10.1016/j.celrep.2022.111187)—exhibited significant upregulation, highlighting the robustness of the data. |
| Institute | Princeton University |
| Department | Department of Chemistry |
| Laboratory | Prof. Dr. Joshua Rabinowitz |
| Last Name | Mann |
| First Name | Charlotte Greta |
| Address | Joseph Stelzmann Strasse 26, 50931 Cologne, Germany |
| cmann5@uni-koeln.de | |
| Phone | +49 221 478-84102 |
| Submit Date | 2025-03-20 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | mzXML |
| Analysis Type Detail | LC-MS |
| Release Date | 2025-04-18 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR002385 |
| Project DOI: | doi: 10.21228/M8BR77 |
| Project Title: | Sulfur Amino Acid Restriction Enhances Exercise Capacity in Mice by Boosting Fat Oxidation in Muscle |
| Project Summary: | Dietary restriction of the sulfur-containing amino acids methionine and cysteine (SAAR) improves body composition, enhances insulin sensitivity, and extends lifespan; benefits seen also with endurance exercise. Yet, the impact of SAAR on skeletal muscle remains largely unexplored. Here we demonstrate that one week of SAAR in sedentary, young, male mice increases endurance exercise capacity. Indirect calorimetry showed that SAAR increased lipid oxidation at rest and delayed the onset of carbohydrate utilization during exercise. Transcriptomic analysis revealed increased expression of genes involved in fatty acid catabolism especially in glycolytic muscle following SAAR. These findings were functionally supported by increased fatty acid circulatory turnover flux and muscle β-oxidation. Reducing lipid uptake from circulation through endothelial cell (EC)-specific CD36 deletion attenuated the running phenotype. Mechanistically, VEGF-signaling inhibition prevented exercise increases following SAAR, without affecting angiogenesis, implicating noncanonical VEGF signaling and EC CD36-dependent fatty acid transport in regulating exercise capacity by influencing muscle substrate availability. |
| Institute: | Princeton University |
| Department: | Department of Chemistry |
| Laboratory: | Prof. Dr. Joshua Rabinowitz |
| Last Name: | Mann |
| First Name: | Charlotte G. |
| Address: | Joseph Stelzmann Strasse 26, 50931 Cologne, Germany |
| Email: | cmann5@uni-koeln.de |
| Phone: | +49 221 478-84102 |
Subject:
| Subject ID: | SU003947 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Genotype Strain: | c57bl/6j |
| Age Or Age Range: | 14 weeks |
| Gender: | Male |
| Animal Feed: | Con versus SAAR diet |
Factors:
Subject type: Mammal; Subject species: Mus musculus (Factor headings shown in green)
| mb_sample_id | local_sample_id | Sample source | Diet |
|---|---|---|---|
| SA417578 | b1 | blank | - |
| SA417579 | b2 | blank | - |
| SA417580 | B3 | blank | - |
| SA417554 | 7 | Mouse EDL muscle | Con |
| SA417555 | 11 | Mouse EDL muscle | Con |
| SA417556 | 9 | Mouse EDL muscle | Con |
| SA417557 | 1 | Mouse EDL muscle | Con |
| SA417558 | 5 | Mouse EDL muscle | Con |
| SA417559 | 3 | Mouse EDL muscle | Con |
| SA417560 | 6 | Mouse EDL muscle | SAAR |
| SA417561 | 8 | Mouse EDL muscle | SAAR |
| SA417562 | 4 | Mouse EDL muscle | SAAR |
| SA417563 | 10 | Mouse EDL muscle | SAAR |
| SA417564 | 12 | Mouse EDL muscle | SAAR |
| SA417565 | 2 | Mouse EDL muscle | SAAR |
| SA417566 | 23 | Mouse Soleus muscle | Con |
| SA417567 | 21 | Mouse Soleus muscle | Con |
| SA417568 | 19 | Mouse Soleus muscle | Con |
| SA417569 | 17 | Mouse Soleus muscle | Con |
| SA417570 | 15 | Mouse Soleus muscle | Con |
| SA417571 | 13 | Mouse Soleus muscle | Con |
| SA417572 | 18 | Mouse Soleus muscle | SAAR |
| SA417573 | 16 | Mouse Soleus muscle | SAAR |
| SA417574 | 20 | Mouse Soleus muscle | SAAR |
| SA417575 | 14 | Mouse Soleus muscle | SAAR |
| SA417576 | 22 | Mouse Soleus muscle | SAAR |
| SA417577 | 24 | Mouse Soleus muscle | SAAR |
| Showing results 1 to 27 of 27 |
Collection:
| Collection ID: | CO003940 |
| Collection Summary: | Tissues were collected and snap-frozen in liquid nitrogen. |
| Sample Type: | Muscle |
Treatment:
| Treatment ID: | TR003956 |
| Treatment Summary: | Wild type (WT) C57BL/6J were purchased from Charles River (Freiburg im Breisgau, Germany). Experimental diets were based on Research Diets D12450J with approximately 18% of calories from protein, 10% from fat and 72% from carbohydrates. SAAR diets containing 1.15g methionine (M)/kg food and lacking cysteine (C) (Miller et al., 2005, DOI: 10.1111/j.1474-9726.2005.00152.x ) in the context of a 17% protein/ 73% carbohydrate calorie diet were provided Ad Libitum (AL). Food intake was monitored daily during experiments. The Research Diets product number for the control diet is A17101101 and for SAAR diet is A17101103. |
Sample Preparation:
| Sampleprep ID: | SP003953 |
| Sampleprep Summary: | Metabolite extraction of tissues Frozen tissue pieces were pulverized using a Cryomill (Retsch) at cryogenic temperature. Ground tissue was weighed (10–20 mg) and transferred into a precooled tube for extraction. Soluble metabolites extraction was done by adding −20 °C 40:40:20 methanol:acetonitrile:water to the resulting powder (40 μl solvent per mg tissue). Samples were vortexed for 10 seconds, cooled at 4°C (on wet ice) for 20 minutes and then centrifuged at 4 °C at 20,000 x g for 30 minutes. Supernatant was transferred to LC–MS vials for analysis. |
Combined analysis:
| Analysis ID | AN006269 | AN006270 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | HILIC | HILIC |
| Chromatography system | Thermo Dionex | Thermo Dionex |
| Column | Waters XBridge BEH Amide (100 x 2.1mm,2.5um) | Waters XBridge BEH Amide (100 x 2.1mm,2.5um) |
| MS Type | ESI | ESI |
| MS instrument type | Orbitrap | Orbitrap |
| MS instrument name | Thermo Orbitrap Exploris 480 | Thermo Orbitrap Exploris 480 |
| Ion Mode | POSITIVE | NEGATIVE |
| Units | ion count | ion count |
Chromatography:
| Chromatography ID: | CH004756 |
| Chromatography Summary: | Solvent A was maintained at pH 9.4 |
| Instrument Name: | Thermo Dionex |
| Column Name: | Waters XBridge BEH Amide (100 x 2.1mm,2.5um) |
| Column Temperature: | 25°C |
| Flow Gradient: | 0 minutes, 85% B; 2 minutes, 85% B; 3 minutes, 80% B; 5 minutes, 80% B; 6 minutes, 75% B; 7 minutes, 75% B; 8 minutes, 70% B; 9 minutes, 70% B; 10 minutes, 50% B; 12 minutes, 50% B; 13 minutes, 25% B; 16 minutes, 25% B; 18 minutes, 0% B; 23 minutes, 0% B; 24 minutes, 85% B; 30 minutes, 85% B |
| Flow Rate: | 150 μl min−1 |
| Solvent A: | 95% water/5% acetonitrile; 20 mM ammonium acetate; 20 mM ammonium hydroxide |
| Solvent B: | 100% acetonitrile |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS005971 |
| Analysis ID: | AN006269 |
| Instrument Name: | Thermo Orbitrap Exploris 480 |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | MS full scans were in negative or positive ion mode with a resolution of 140,000 at m/z 200 and scan range of 70–1,000 m/z. The automatic gain control (AGC) target was 1 × 106. LC-MS peak files were analyzed and visualized with El-MAVEN (Elucidata) using 5 ppm ion extraction window, minimum peak intensity of 1 x 105 ions, and minimum signal to background blank ratio of 2 |
| Ion Mode: | POSITIVE |
| MS ID: | MS005972 |
| Analysis ID: | AN006270 |
| Instrument Name: | Thermo Orbitrap Exploris 480 |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | MS full scans were in negative or positive ion mode with a resolution of 140,000 at m/z 200 and scan range of 70–1,000 m/z. The automatic gain control (AGC) target was 1 × 106. LC-MS peak files were analyzed and visualized with El-MAVEN |
| Ion Mode: | NEGATIVE |
