Summary of Study ST000468

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 PR000360. The data can be accessed directly via it's Project DOI: 10.21228/M8HK59 This work is supported by NIH grant, U2C- DK119886.

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Study IDST000468
Study TitleEnterococcus faecalis nucleotide profiles following mupirocin or decoyinine
Study TypeExposure to mupirocin or decoyinine
Study SummaryTriplicate samples of E. faecalis OG1RF will be grown in FMC-AUG to OD600 = 0.25. The cultures will be split and exposed to DMSO, mupirocin (0.01mg/mL) or decoyinine (0.1mg/mL) for 15 minutes. 6E9 cells will be harvested by fast filtration, scraped off of membrane into ice-cold PBS, and pelleted by centrifugation, and stored at -80C until shipment.
Institute
University of Florida
DepartmentSECIM
LaboratoryLemos-Abranches
Last NameKajfasz
First NameJessica
Address1395 Center Drive, PO Box 100424 Gainesville, FL 32610
Emailjkajfasz@dental.ufl.edu
Phone352-273-6675
Submit Date2016-09-08
Num Groups3
Total Subjects9
Raw Data File Type(s)d
Analysis Type DetailLC-MS
Release Date2017-10-03
Release Version1
Jessica Kajfasz Jessica Kajfasz
https://dx.doi.org/10.21228/M8HK59
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR000360
Project DOI:doi: 10.21228/M8HK59
Project Title:Enterococcus faecalis nucleotie profiles
Project Summary:Determination of metabolite profile of E. faecalis OG1RF control as compared to exposure to mupirocin or decoyinine
Institute:University of Florida
Department:Oral Biology
Laboratory:Oral Biology Lemos-Abranches
Last Name:Lemos
First Name:Jose
Address:1395 Center Drive, PO Box 100424 Gainesville, FL 32610
Email:jlemos@dental.ufl.edu
Phone:352-273-8843
Funding Source:UF start-up funds

Subject:

Subject ID:SU000489
Subject Type:Bacterial cells
Subject Species:Enterococcus faecalis
Taxonomy ID:1351
Genotype Strain:OG1RF
Species Group:Microorganism

Factors:

Subject type: Bacterial cells; Subject species: Enterococcus faecalis (Factor headings shown in green)

mb_sample_id local_sample_id Exposure
SA023657OG1RF_1 DecoyDecoyinine, 0.1mg/mL
SA023658OG1RF_2 DecoyDecoyinine, 0.1mg/mL
SA023659OG1RF_3 DecoyDecoyinine, 0.1mg/mL
SA023654OG1RF_1 DMSODMSO
SA023655OG1RF_2 DMSODMSO
SA023656OG1RF_3 DMSODMSO
SA023660OG1RF_2 MupMupirocin, 0.01mg/mL
SA023661OG1RF_1 MupMupirocin, 0.01mg/mL
SA023662OG1RF_3 MupMupirocin, 0.01mg/mL
Showing results 1 to 9 of 9

Collection:

Collection ID:CO000483
Collection Summary:After the 15 minute treatment interval, 20mL sample will be harvested by filtration onto 0.45uM nitrocellulose membrane. The cells will be scraped off of the membrane into ice-cold PBS (~1mL), centrifuged for 45 seconds at 12K RPM, liquid aspirated and stored at -80C until sending for analysis.
Sample Type:Bacterial cells
Collection Method:Fast filtration
Volumeoramount Collected:20ml
Storage Conditions:-80C

Treatment:

Treatment ID:TR000503
Treatment Summary:Triplicate cultures of E. faecalis OG1RF will be grown in FMC-AUG to OD600 = 0.25. The cultures will be split into 3 equal volumes of 30mL which will be exposed to 100uL DMSO, 0.01mg/mL mupirocin, or 0.1mg/mL decoyinine for 15 minutes. At this point, 10mL sample will be removed for protein analysis. The remaining 20mL will be harvested by fast filtration for analysis.|Triplicate cultures of E. faecalis OG1RF will be grown in FMC-AUG to OD600 = 0.25. The cultures will be split into 3 equal volumes of 30mL which will be exposed to 100uL DMSO, 0.01mg/mL mupirocin, or 0.1mg/mL decoyinine for 15 minutes. At this point, 10mL sample will be removed for protein analysis. The remaining 20mL will be harvested by fast filtration for analysis.|Triplicate cultures of E. faecalis OG1RF will be grown in FMC-AUG to OD600 = 0.25. The cultures will be split into 3 equal volumes of 30mL which will be exposed to 100uL DMSO, 0.01mg/mL mupirocin, or 0.1mg/mL decoyinine for 15 minutes. At this point, 10mL sample will be removed for protein analysis. The remaining 20mL will be harvested by fast filtration for analysis.
Treatment Compound:DMSO|Mupirocin|Decoyinine
Treatment Dose:100uL|0.01mg/mL|0.1mg/mL
Treatment Doseduration:15 min|15 min|15 min
Treatment Vehicle:DMSO|DMSO|DMSO

Sample Preparation:

Sampleprep ID:SP000496
Sampleprep Summary:NAD-ATP
Sampleprep Protocol Filename:SB_NAD-ATP_Assay_Agilent.pdf

Combined analysis:

Analysis ID AN000731
Analysis type MS
Chromatography type Reversed phase
Chromatography system Agilent 1290 Infinity
Column Thermo Hypercarb (50 x 2.1mm, 3um)
MS Type ESI
MS instrument type Triple quadrupole
MS instrument name Agilent 6490 QQQ
Ion Mode POSITIVE
Units nmol/mg protein

Chromatography:

Chromatography ID:CH000524
Chromatography Summary:NAD-ATP
Methods Filename:SB_NAD-ATP_Assay_Agilent.pdf
Instrument Name:Agilent 1290 Infinity
Column Name:Thermo Hypercarb (50 x 2.1mm, 3um)
Chromatography Type:Reversed phase

MS:

MS ID:MS000648
Analysis ID:AN000731
Instrument Name:Agilent 6490 QQQ
Instrument Type:Triple quadrupole
MS Type:ESI
Ion Mode:POSITIVE
Analysis Protocol File:SB_NAD-ATP_Assay_Agilent.pdf
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