Summary of Study ST004013
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 PR002501. The data can be accessed directly via it's Project DOI: 10.21228/M8C55X 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 | ST004013 |
| Study Title | Aqueous solubility of MMTC/57E in phosphate-buffered saline at room temperature. |
| Study Summary | The study was designed to quantify the absolute concentration of soluble 57E/MMTC present in PBS following a mock activity assay in tissue culture plates. |
| Institute | University of British Columbia |
| Department | Biochemistry & Molecular Biology |
| Laboratory | Parker laboratory |
| Last Name | Parker |
| First Name | Seth |
| Address | 950 W 28th Ave, Vancouver, British Columbia, V6H 0B3, Canada |
| seth.parker@bcchr.ca | |
| Phone | 6048753121 |
| Submit Date | 2025-06-19 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | mzML |
| Analysis Type Detail | LC-MS |
| Release Date | 2025-07-30 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Project:
| Project ID: | PR002501 |
| Project DOI: | doi: 10.21228/M8C55X |
| Project Title: | Metabolomics analysis of SNAT2-deficient cells: implications for the discovery of selective transporter inhibitors |
| Project Type: | Manuscript |
| Project Summary: | Amino acid uptake by the solute carrier family of transporter proteins is critical to support cell metabolism, and inhibition of transporter activity represents a tractable strategy to restrict nutrient availability to cancer cells. A small molecule inhibitor of the sodium-coupled neutral amino acid transporter 2 (SNAT2), 3-(N-methyl(4-methylphenyl)sulfonamido)-N-(2-trifluoromethylbenzyl)thiophene-2-carboxamide (MMTC/57E), was recently identified and was shown to inhibit cell proliferation when combined with glucose transport inhibitors in breast and pancreatic cancer cell lines. In this study, we use mass spectrometry-based metabolomics and establish cell-based assays for the SNAT2 transporter. We show that SNAT2 knockout cells have significant defects in amino acid availability. Using our established assays, we fail to observe that MMTC/57E inhibits SNAT2 activity likely due to its poor solubility. |
| Institute: | University of British Columbia |
| Department: | Biochemistry & Molecular Biology |
| Laboratory: | Parker laboratory |
| Last Name: | Parker |
| First Name: | Seth |
| Address: | 950 W 28th Ave, Vancouver, British Columbia, V6H 0B3, Canada |
| Email: | seth.parker@bcchr.ca |
| Phone: | 6048753121 |
Subject:
| Subject ID: | SU004151 |
| Subject Type: | Synthetic sample |
Factors:
Subject type: Synthetic sample; Subject species: - (Factor headings shown in green)
| mb_sample_id | local_sample_id | Factor | Sample source |
|---|---|---|---|
| SA461794 | 57E_std5_r2 | none | Standard Curve |
| SA461795 | 57E_std1_r1 | none | Standard Curve |
| SA461796 | 57E_std1_r2 | none | Standard Curve |
| SA461797 | 57E_std2_r1 | none | Standard Curve |
| SA461798 | 57E_std2_r2 | none | Standard Curve |
| SA461799 | 57E_std3_r1 | none | Standard Curve |
| SA461800 | 57E_std3_r2 | none | Standard Curve |
| SA461801 | 57E_std4_r2 | none | Standard Curve |
| SA461802 | 57E_std5_r1 | none | Standard Curve |
| SA461803 | 57E_std4_r1 | none | Standard Curve |
| SA461804 | 57E_std6_r1 | none | Standard Curve |
| SA461805 | 57E_std9_r1 | none | Standard Curve |
| SA461806 | 57E_std6_r2 | none | Standard Curve |
| SA461807 | 57E_std10_r1 | none | Standard Curve |
| SA461808 | 57E_std9_r2 | none | Standard Curve |
| SA461809 | 57E_std10_r2 | none | Standard Curve |
| SA461810 | 57E_std8_r2 | none | Standard Curve |
| SA461811 | 57E_std8_r1 | none | Standard Curve |
| SA461812 | 57E_std7_r2 | none | Standard Curve |
| SA461813 | 57E_std7_r1 | none | Standard Curve |
| SA461814 | 57E_solu_5DMSO_10uM_filt_1 | post-filter | 57E Solubility |
| SA461815 | 57E_solu_5DMSO_40uM_filt_3 | post-filter | 57E Solubility |
| SA461816 | 57E_solu_5DMSO_40uM_filt_2 | post-filter | 57E Solubility |
| SA461817 | 57E_solu_5DMSO_40uM_filt_1 | post-filter | 57E Solubility |
| SA461818 | 57E_solu_5DMSO_20uM_filt_3 | post-filter | 57E Solubility |
| SA461819 | 57E_solu_5DMSO_20uM_filt_2 | post-filter | 57E Solubility |
| SA461820 | 57E_solu_5DMSO_20uM_filt_1 | post-filter | 57E Solubility |
| SA461821 | 57E_10microM_filt_r8 | post-filter | 57E Solubility |
| SA461822 | 57E_solu_5DMSO_10uM_filt_3 | post-filter | 57E Solubility |
| SA461823 | 57E_solu_5DMSO_10uM_filt_2 | post-filter | 57E Solubility |
| SA461824 | 57E_10microM_filt_r7 | post-filter | 57E Solubility |
| SA461825 | 57E_10microM_filt_r9 | post-filter | 57E Solubility |
| SA461826 | 57E_20microM_filt_r7 | post-filter | 57E Solubility |
| SA461827 | 57E_40microM_filt_r7 | post-filter | 57E Solubility |
| SA461828 | 57E_40microM_filt_r8 | post-filter | 57E Solubility |
| SA461829 | 57E_40microM_filt_r9 | post-filter | 57E Solubility |
| SA461830 | 57E_20microM_filt_r8 | post-filter | 57E Solubility |
| SA461831 | 57E_20microM_filt_r9 | post-filter | 57E Solubility |
| SA461832 | PBS_filt_r7 | post-filter | Control (PBS) |
| SA461833 | 57E_solu_5DMSO_PBS_filt_2 | post-filter | Control (PBS) |
| SA461834 | 57E_solu_5DMSO_PBS_filt_3 | post-filter | Control (PBS) |
| SA461835 | 57E_solu_5DMSO_PBS_filt_1 | post-filter | Control (PBS) |
| SA461836 | PBS_filt_r9 | post-filter | Control (PBS) |
| SA461837 | PBS_filt_r8 | post-filter | Control (PBS) |
| SA461838 | 57E_solu_5DMSO_40uM_init_1 | post-shake | 57E Solubility |
| SA461839 | 57E_40microM_r3 | post-shake | 57E Solubility |
| SA461840 | 57E_40microM_r4 | post-shake | 57E Solubility |
| SA461841 | 57E_40microM_r2 | post-shake | 57E Solubility |
| SA461842 | 57E_10microM_r6 | post-shake | 57E Solubility |
| SA461843 | 57E_10microM_r5 | post-shake | 57E Solubility |
| SA461844 | 57E_10microM_r4 | post-shake | 57E Solubility |
| SA461845 | 57E_solu_5DMSO_40uM_init_3 | post-shake | 57E Solubility |
| SA461846 | 57E_solu_5DMSO_40uM_init_2 | post-shake | 57E Solubility |
| SA461847 | 57E_solu_0pt1DMSO_20uM_init_1 | post-shake | 57E Solubility |
| SA461848 | 57E_solu_5DMSO_20uM_init_2 | post-shake | 57E Solubility |
| SA461849 | 57E_20microM_r1 | post-shake | 57E Solubility |
| SA461850 | 57E_20microM_r2 | post-shake | 57E Solubility |
| SA461851 | 57E_20microM_r3 | post-shake | 57E Solubility |
| SA461852 | 57E_40microM_r1 | post-shake | 57E Solubility |
| SA461853 | 57E_20microM_r6 | post-shake | 57E Solubility |
| SA461854 | 57E_solu_5DMSO_20uM_init_3 | post-shake | 57E Solubility |
| SA461855 | 57E_40microM_r5 | post-shake | 57E Solubility |
| SA461856 | 57E_solu_5DMSO_20uM_init_1 | post-shake | 57E Solubility |
| SA461857 | 57E_solu_0pt1DMSO_10uM_init_3 | post-shake | 57E Solubility |
| SA461858 | 57E_solu_0pt1DMSO_20uM_init_2 | post-shake | 57E Solubility |
| SA461859 | 57E_solu_0pt1DMSO_20uM_init_3 | post-shake | 57E Solubility |
| SA461860 | 57E_solu_0pt1DMSO_40uM_init_1 | post-shake | 57E Solubility |
| SA461861 | 57E_solu_0pt1DMSO_40uM_init_2 | post-shake | 57E Solubility |
| SA461862 | 57E_solu_0pt1DMSO_40uM_init_3 | post-shake | 57E Solubility |
| SA461863 | 57E_20microM_r4 | post-shake | 57E Solubility |
| SA461864 | 57E_10microM_r1 | post-shake | 57E Solubility |
| SA461865 | 57E_20microM_r5 | post-shake | 57E Solubility |
| SA461866 | 57E_solu_5DMSO_10uM_init_1 | post-shake | 57E Solubility |
| SA461867 | 57E_10microM_r2 | post-shake | 57E Solubility |
| SA461868 | 57E_40microM_r6 | post-shake | 57E Solubility |
| SA461869 | 57E_solu_0pt1DMSO_10uM_init_1 | post-shake | 57E Solubility |
| SA461870 | 57E_solu_5DMSO_10uM_init_3 | post-shake | 57E Solubility |
| SA461871 | 57E_10microM_r3 | post-shake | 57E Solubility |
| SA461872 | 57E_solu_0pt1DMSO_10uM_init_2 | post-shake | 57E Solubility |
| SA461873 | 57E_solu_5DMSO_10uM_init_2 | post-shake | 57E Solubility |
| SA461874 | PBS_r6 | post-shake | Control (PBS) |
| SA461875 | PBS_r5 | post-shake | Control (PBS) |
| SA461876 | PBS_r4 | post-shake | Control (PBS) |
| SA461877 | 57E_solu_5DMSO_PBS_init_3 | post-shake | Control (PBS) |
| SA461878 | 57E_solu_5DMSO_PBS_init_2 | post-shake | Control (PBS) |
| SA461879 | 57E_solu_5DMSO_PBS_init_1 | post-shake | Control (PBS) |
| SA461880 | 57E_solu_0pt1DMSO_PBS_init_1 | post-shake | Control (PBS) |
| SA461881 | 57E_solu_0pt1DMSO_PBS_init_2 | post-shake | Control (PBS) |
| SA461882 | Blank_PBS_r3 | post-shake | Control (PBS) |
| SA461883 | Blank_PBS_r2 | post-shake | Control (PBS) |
| SA461884 | Blank_PBS_r1 | post-shake | Control (PBS) |
| SA461885 | 57E_solu_0pt1DMSO_PBS_init_3 | post-shake | Control (PBS) |
| SA461886 | 57E_solu_0pt1DMSO_10uM_ps_3 | post-spin | 57E Solubility |
| SA461887 | 57E_solu_5DMSO_10uM_ps_2 | post-spin | 57E Solubility |
| SA461888 | 57E_solu_5DMSO_10uM_ps_1 | post-spin | 57E Solubility |
| SA461889 | 57E_solu_0pt1DMSO_20uM_ps_1 | post-spin | 57E Solubility |
| SA461890 | 57E_solu_0pt1DMSO_20uM_ps_2 | post-spin | 57E Solubility |
| SA461891 | 57E_solu_0pt1DMSO_20uM_ps_3 | post-spin | 57E Solubility |
| SA461892 | 57E_solu_0pt1DMSO_40uM_ps_1 | post-spin | 57E Solubility |
| SA461893 | 57E_solu_0pt1DMSO_40uM_ps_2 | post-spin | 57E Solubility |
Collection:
| Collection ID: | CO004144 |
| Collection Summary: | A 10 mM stock of MMTC/57E in DMSO was diluted into 1 mL of PBS at a final DMSO concentration of either 0.1% or 5% at either 10, 20 or 40 µM. The samples were vortexed at ~500 rpm in a polystyrene 12-well tissue culture plate for 30 minutes at 20°C. 10 µL was transferred to a glass vial containing 90 µL of 80% acetonitrile in water (post-shake sample). The remaining volume was transferred to a 1.5 mL tube and centrifuged at 21,300 x g for 5 minutes at 20°C. 10 µL of the supernatant was transferred to a glass vial containing 90 uL of 80% acetonitrile in water (post-spin sample). 500 µL of the supernatant was transferred to a syringe and filtered through a 0.22 micron low-bind polyethersulfone (PES) filter. 10 µL of the filtrate was added to a glass vial containing 90 µL of 80% acetonitrile in water (post-filter sample). All samples were analyzed by LCMS, and the absolute concentration was quantified using a standard curve generated by preparing a 2.5 µM stock of 57E in 80% acetonitrile in water and a 10-point serial dilution. |
| Sample Type: | Media |
Treatment:
| Treatment ID: | TR004160 |
| Treatment Summary: | No treatment. |
Sample Preparation:
| Sampleprep ID: | SP004157 |
| Sampleprep Summary: | Samples were prepared directly in 80% acetonitrile in water. 2 µL of each sample was injected. The final concentrations calculated taking into account the 10-fold dilution for the solubility samples. |
Chromatography:
| Chromatography ID: | CH005028 |
| Chromatography Comments: | Only the first 4 minutes of the LCMS run were included, as 57E elutes at 2.8 minutes. |
| Instrument Name: | Thermo Vanquish |
| Column Name: | Merck SeQuant ZIC-pHILIC (150 x 2.1 mm, 5 µm) |
| Column Temperature: | 25°C |
| Flow Gradient: | 80-20%B (0-30 min), 20-20%B (30-40 minute), and 20-80%B (40-40.5 minute); the LC column was re-equilibrated using 80-80%B from 40.5-52 minute before subsequent injections |
| Flow Rate: | 100 µL/min |
| Solvent A: | 100% Water; 10 mM Ammonium Carbonate, pH 9.0 |
| Solvent B: | 100% Acetonitrile |
| Chromatography Type: | HILIC |
Analysis:
| Analysis ID: | AN006618 |
| Analysis Type: | MS |
| Chromatography ID: | CH005028 |
| Num Factors: | 7 |
| Num Metabolites: | 1 |
| Units: | ion counts |
