Summary of Study ST003068
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 PR001913. The data can be accessed directly via it's Project DOI: 10.21228/M8FH9H 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 | ST003068 |
Study Title | Attenuation of Helicobacter pylori VacA toxin-induced cell death by modulation of intracellular taurine metabolism _ Study #2 |
Study Type | untargeted metabolomics analysis |
Study Summary | AGS cells treated with wild-type VacA have metabolic profiles different from those of AGS cells treated with mutant forms of VacA. Most VacA-induced cellular alterations are attributed to VacA’s ability to form channels within host cell membranes. To determine if the observed VacA-induced metabolic alterations were dependent on VacA’s ability to form membrane channels, we treated AGS cells with the wild-type s1m1 wild-type (WT) toxin (which has robust channel-forming activity), as well as two mutant forms of the toxin, Delta_6-27 or s2m1. VacA Delta_6-27 has a deletion in the VacA amino-terminal hydrophobic region and is defective in channel formation. In the s2m1 mutant toxin, the active s1 isotype sequence of the WT toxin is replaced with the less active s2 sequence, resulting in markedly diminished channel-forming activity. AGS cells treated with the Delta_6-27 or s2m1 mutant VacA did not undergo vacuolation, while AGS cells treated with WT VacA experienced robust vacuolation, consistent with previously published results. |
Institute | Vanderbilt University |
Department | Chemistry |
Laboratory | Center for Innovative Technology |
Last Name | CODREANU |
First Name | SIMONA |
Address | 1234 STEVENSON CENTER LANE |
SIMONA.CODREANU@VANDERBILT.EDU | |
Phone | 6158758422 |
Submit Date | 2024-02-05 |
Raw Data Available | Yes |
Raw Data File Type(s) | raw(Thermo) |
Analysis Type Detail | LC-MS |
Release Date | 2024-06-12 |
Release Version | 1 |
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Collection:
Collection ID: | CO003176 |
Collection Summary: | AGS cells or AZ-521 cells were cultured in Roswell Park Memorial Institute (RPMI) 1640 medium containing 10% fetal bovine serum, or minimal essential medium (MEM) containing 10% fetal bovine serum and 5% nonessential amino acids, respectively. AGS cells were seeded at 2x104 cells/well into 96-well plates and incubated overnight. Cultured cells were then incubated with 20 ug/mL purified VacA [activated with by addition of HCl to a pH of 3] in medium supplemented with 5 mM NH4Cl. Following intoxication, the media was removed, and cells were washed with PBS. Cells were detached by incubation with trypsin for 5 minutes and collected via centrifugation at 4°C at 1,000 rpm for 4 minutes. Trypsin was removed, cells were once again washed with PBS, and the cells were then flash-frozen in liquid nitrogen and stored at -70°C. |
Collection Protocol Filename: | Cell_Culture_Methods.pdf |
Sample Type: | Cultured cells |
Storage Conditions: | -80℃ |