Summary of Study ST001904
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 PR001199. The data can be accessed directly via it's Project DOI: 10.21228/M8QQ5J 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 | ST001904 |
| Study Title | Lipidomics analysis of outer membrane vesicles and elucidation of the ceramide phosphoinositol biosynthetic pathway in Bacteroides thetaiotaomicron |
| Study Type | Lipidic profile in wild-type and mutant strains |
| Study Summary | In this work, we characterized the lipid composition of membranes and OMV from Bacteroides thetaiotaomicron VPI-5482. LC-MS analysis indicate that OMV carry sphingolipids, glycerophospholipids and serine-dipeptide lipids. Sphingolipid species represent more than 50% of the total lipid content of OMV. The most abundant sphingolipids in OMV are ceramide phosphoethanolamine (CerPE) and ceramide phosphoinositol (CerPI). Bioinformatic analysis allowed the identification of the BT1522-1526 operon putatively involved in CerPI synthesis. Mutagenesis studies revealed BT1522-1526 are essential for synthesis of PI and CerPI, confirming the role of this operon in biosynthesis of CerPI. BT1522-1526 mutant strains lacking CerPI produced OMV that were indistinguishable from the wild-type strain, indicating that CerPI sphingolipid species are not involved in OMV biogenesis. Bacteroides sphingolipids are thought to modulate host-commensal interactions, and based on our data, we propose that OMV could act as long distance delivery vehicles for these molecules. |
| Institute | Washington University in St. Louis |
| Department | Molecular Microbiology |
| Laboratory | Feldman lab |
| Last Name | Sartorio |
| First Name | Mariana |
| Address | 660 S Euclid avenue, campus box 8230, 63110 |
| mgsartorio@wustl.edu | |
| Phone | 3147474477 |
| Submit Date | 2021-06-22 |
| Analysis Type Detail | LC-MS |
| Release Date | 2021-08-30 |
| Release Version | 1 |
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Collection:
| Collection ID: | CO001975 |
| Collection Summary: | B. thetaiotaomicron strains (wild-type and ΔBT1522-BT1526 mutants) were grown overnight in an anaerobic chamber (Coy Laboratories) using an atmosphere of 10% H2, 5% CO2, 85% N2. For liquid growth, Brain heart infusion (BHI) supplemented with hemin and vitamin K3 was used. Cultures were then subjected to subcellular fractionation to obtain total membranes and outer membrane vesicles (OMV) preparation. OMV preparations: OMV were purified by ultracentrifugation of filtered spent media from 150 ml of liquid culture as described previously (1). OMV preparations were resuspended in PBS before lipid analyses. Protein content was quantified using a DC protein assay kit (Bio-Rad). Fractions were aliquoted and stored at -80°C until analyzed. Membrane preparations: Total membrane preparations were performed by cell lysis and ultracentrifugation as previously described (1). Total membranes from 150 ml of liquid culture were resuspended in PBS using a 2-ml glass tissue grinder with a polytetrafluoroethylene (PTFE) pestle (VWR). Protein content was quantified using a DC protein assay kit (Bio-Rad). Fractions were stored aliquoted and stored at -80°C until analyzed. References: 1. Elhenawy W, Debelyy MO, Feldman MF. Preferential packing of acidic glycosidases and proteases into Bacteroides outer membrane vesicles. mBio. 2014;5(2):e00909-14. |
| Sample Type: | Bacterial cells |
