Summary of Study ST003401

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 PR002107. The data can be accessed directly via it's Project DOI: 10.21228/M88C05 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.

Perform statistical analysis  |  Show all samples  |  Show named metabolites  |  Download named metabolite data  
Download mwTab file (text)   |  Download mwTab file(JSON)   |  Download data files (Contains raw data)
Study IDST003401
Study TitleThe double-edged role of FASII regulator FabT in Streptococcus pyogenes infection - Lipidomics
Study SummaryA FabT mutation impacts the membrane fatty acid (FA) composition. Lipids are composed of FA. The effects of the FabT mutation on membrane FA composition should also alter phospholipid metabolism. WT and mFabT strains were grown as 200 ml cultures in THY or THY-Tween until OD600 = 0.4 - 0.5. Lipid extractions and identifications were performed as described 1-4. Lipid separation was realized by normal phase HPLC (U3000 ThermoFisher Scientific) using a Inertsil Si 5μm column (150 x 2.1 mm I.D.) from GL Sciences Inc (Tokyo, Japan). Lipids were quantified using a Corona-CAD Ultra and identified by mass-spectrometry negative ionization and MS2/MS3 fragmentations (LTQ-Orbitrap Velos Pro). The adducts observed for molecular ions were: CH3COO- for MGDG, CH3COO- and H- for DGDG, H- for PG and CL. All raw data from MS/MS fragmentations can now be accessed in MZML open source format. This analysis was repeated on 3 independent experiments. 1-Kenanian, G. et al. Permissive Fatty Acid Incorporation Promotes Staphylococcal Adaptation to FASII Antibiotics in Host Environments. Cell Rep 29, 3974-3982 e3974, doi:10.1016/j.celrep.2019.11.071 (2019). 2-Abreu, S., Solgadi, A. & Chaminade, P. Optimization of normal phase chromatographic conditions for lipid analysis and comparison of associated detection techniques. J Chromatogr A 1514, 54-71, doi:10.1016/j.chroma.2017.07.063 (2017). 3-Bligh, E. G. & Dyer, W. J. A rapid method of total lipid extraction and purification. Can J Biochem Physiol 37, 911-917, doi:10.1139/o59-099 (1959). 4-Thedieck, K. et al. The MprF protein is required for lysinylation of phospholipids in listerial membranes and confers resistance to cationic antimicrobial peptides (CAMPs) on Listeria monocytogenes. Mol Microbiol 62, 1325-1339, doi:10.1111/j.1365-2958.2006.05452.x (2006).
Institute
INSERM
DepartmentU1016
LaboratoryInstitut Cochin - Bacteria and perinatality team
Last NameLambert
First NameClara
Address24 rue Méchain 75014 Paris
Emailclara.lambert@ibcp.fr
Phone+33134652168
Submit Date2024-08-02
Raw Data AvailableYes
Raw Data File Type(s)mzML, raw(Thermo)
Analysis Type DetailAPCI
Release Date2024-08-19
Release Version1
Clara Lambert Clara Lambert
https://dx.doi.org/10.21228/M88C05
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

Select appropriate tab below to view additional metadata details:

Project:

Project ID:PR002107
Project DOI:doi: 10.21228/M88C05
Project Title:The double-edged role of FASII regulator FabT in Streptococcus pyogenes infection
Project Type:MS quantitative analysis
Project Summary:In Streptococcus pyogenes, the type II fatty acid (FA) synthesis pathway FASII is feedback-controlled by the FabT repressor. A FabT mutation leads to FASII dysregulation, FAs membrane modification and bacterial growth defect in the presence of eukaryotic cells and their supernatant. We try to understand the consequences on bacteria metabolism.
Institute:INSERM
Department:U1016
Laboratory:Institut Cochin - Bacteria and perinatality team
Last Name:Lambert
First Name:Clara
Address:24 rue Méchain 75014 Paris
Email:clara.lambert@ibcp.fr
Phone:+33134652168
Publications:The double-edged role of FASII regulator FabT in Streptococcus pyogenes infection

Subject:

Subject ID:SU003526
Subject Type:Bacteria
Subject Species:Streptococcus pyogenes
Genotype Strain:M28PF1
Gender:Not applicable

Factors:

Subject type: Bacteria; Subject species: Streptococcus pyogenes (Factor headings shown in green)

mb_sample_id local_sample_id Sample source Treatment
SA371867mFabT_THY_3Bacteria THY
SA371868WT_THY_2Bacteria THY
SA371869mFabT_THY_4Bacteria THY
SA371870WT_THY_1Bacteria THY
SA371871mFabT_THY_2Bacteria THY
SA371872mFabT_THY_1Bacteria THY
SA371873WT_THY_4Bacteria THY
SA371874WT_THY_3Bacteria THY
SA371875WT_THY-Tween_2Bacteria THY-Tween
SA371876WT_THY-Tween_3Bacteria THY-Tween
SA371877WT_THY-Tween_4Bacteria THY-Tween
SA371878mFabT_THY-Tween_1Bacteria THY-Tween
SA371879mFabT_THY-Tween_2Bacteria THY-Tween
SA371880mFabT_THY-Tween_3Bacteria THY-Tween
SA371881mFabT_THY-Tween_4Bacteria THY-Tween
SA371882WT_THY-Tween_1Bacteria THY-Tween
Showing results 1 to 16 of 16

Collection:

Collection ID:CO003519
Collection Summary:Bacteria were grown as 200 ml cultures in THY or THY-Tween until OD600 = 0.4 - 0.5. Lipid extractions and identifications were performed as described 1-4. 1-Kenanian, G. et al. Permissive Fatty Acid Incorporation Promotes Staphylococcal Adaptation to FASII Antibiotics in Host Environments. Cell Rep 29, 3974-3982 e3974, doi:10.1016/j.celrep.2019.11.071 (2019). 2-Abreu, S., Solgadi, A. & Chaminade, P. Optimization of normal phase chromatographic conditions for lipid analysis and comparison of associated detection techniques. J Chromatogr A 1514, 54-71, doi:10.1016/j.chroma.2017.07.063 (2017). 3-Bligh, E. G. & Dyer, W. J. A rapid method of total lipid extraction and purification. Can J Biochem Physiol 37, 911-917, doi:10.1139/o59-099 (1959). 4-Thedieck, K. et al. The MprF protein is required for lysinylation of phospholipids in listerial membranes and confers resistance to cationic antimicrobial peptides (CAMPs) on Listeria monocytogenes. Mol Microbiol 62, 1325-1339, doi:10.1111/j.1365-2958.2006.05452.x (2006).
Sample Type:Bacterial cells

Treatment:

Treatment ID:TR003535
Treatment Summary:WT and mFabT strains were grown as 200 ml cultures in THY or THY-Tween until optical density at 600nm (D600) rich 0.4 - 0.5.

Sample Preparation:

Sampleprep ID:SP003533
Sampleprep Summary:Lipid extractions and identifications were performed as described 1-4. Lipid separation was realized by normal phase HPLC (U3000 ThermoFisher Scientific) using a Inertsil Si 5μm column (150 x 2.1 mm I.D.) from GL Sciences Inc (Tokyo, Japan). Lipids were quantified using a Corona-CAD Ultra and identified by mass-spectrometry negative ionization and MS2/MS3 fragmentations (LTQ-Orbitrap Velos Pro). The adducts observed for molecular ions were: CH3COO- for MGDG, CH3COO- and H- for DGDG, H- for PG and CL. 1-Kenanian, G. et al. Permissive Fatty Acid Incorporation Promotes Staphylococcal Adaptation to FASII Antibiotics in Host Environments. Cell Rep 29, 3974-3982 e3974, doi:10.1016/j.celrep.2019.11.071 (2019). 2-Abreu, S., Solgadi, A. & Chaminade, P. Optimization of normal phase chromatographic conditions for lipid analysis and comparison of associated detection techniques. J Chromatogr A 1514, 54-71, doi:10.1016/j.chroma.2017.07.063 (2017). 3-Bligh, E. G. & Dyer, W. J. A rapid method of total lipid extraction and purification. Can J Biochem Physiol 37, 911-917, doi:10.1139/o59-099 (1959). 4-Thedieck, K. et al. The MprF protein is required for lysinylation of phospholipids in listerial membranes and confers resistance to cationic antimicrobial peptides (CAMPs) on Listeria monocytogenes. Mol Microbiol 62, 1325-1339, doi:10.1111/j.1365-2958.2006.05452.x (2006).

Combined analysis:

Analysis ID AN005582
Analysis type MS
Chromatography type Normal phase
Chromatography system ThermoFisher Ultimate 3000 RSLC
Column Intersil si (150 x 2.1mm, 5um)
MS Type APCI
MS instrument type LTQ-FT
MS instrument name Thermo Velos LTQ Orbitrap
Ion Mode NEGATIVE
Units Relative abundance

Chromatography:

Chromatography ID:CH004242
Instrument Name:ThermoFisher Ultimate 3000 RSLC
Column Name:Intersil si (150 x 2.1mm, 5um)
Column Temperature:40
Flow Gradient:T(min) %A %B %C: 0 100 0 0; 1.5 100 0 0; 1.6 97 3 0; 9 94 6 0; 11 70 30 0; 14 45 55 0; 15 45 55 0; 16 40 55 5; 20 35 55 10; 20.1 33 50 17; 25 38 45 17; 25.1 48 35 17; 30 53 30 17; 40 40 0 60; 40.1 0 100 0; 42 0 100 0; 42.1 50 0 0; 45 50 0 0; 47 100 0 0; 53 100 0 0
Flow Rate:0.8mL/min
Solvent A:100% heptane
Solvent B:66.67% acetone/33.33% ethyl acetate; 0.15% acetic acid
Chromatography Type:Normal phase
Solvent C:85% isopropanol/15% water; 0.043% acetic acid; 0.104 % triethylamine

MS:

MS ID:MS005307
Analysis ID:AN005582
Instrument Name:Thermo Velos LTQ Orbitrap
Instrument Type:LTQ-FT
MS Type:APCI
MS Comments:Fulscan MS, data dependent analysis MS2 (top 2) and MS3 (top 1) Manual asignment compared to commercial standards (Abreu, S., Solgadi, A. & Chaminade, P. Optimization of normal phase chromatographic conditions for lipid analysis and comparison of associated detection techniques. J Chromatogr A 1514, 54-71, doi:10.1016/j.chroma.2017.07.063 (2017).)
Ion Mode:NEGATIVE
  logo