Summary of study ST000154

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 PR000129. The data can be accessed directly via it's Project DOI: 10.21228/M85C7W This work is supported by NIH grant, U2C- DK119886.

See: https://www.metabolomicsworkbench.org/about/howtocite.php

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Study IDST000154
Study TitleUse of Aspartate Dehydrogenase by cancer cells
Study SummaryThe human acute lymphoblastic leukemia cell line (Jurkat) was used to isolate a clones based on Noxa expression. JP6 is low Noxa, JP3 is high Noxa. Furthermore, a Noxa expressing plasmid was tranfected and stable clones were selected for a Noxa high model (N5). 10E6 cells were starved of glucose (A) or glutamine (B&C) for 3 hours and then fed 13C 1,2 glucose (A), 15N alpha nitrogen glutamine (B) or 15N amide nitrogen glutamine (C) for 24 hours. Cells were washed 1X in ice cold PBS and resuspended in -20C methanol. Quenched cells were snap frozen and stored at -80. For experiment A we would like to observe the contribution of labeled glucose into the synthesis of amino acids, specifically glycine and serine. For experiment B we are most interested in the nitrogen incorporation into aspartate. SCB edits: This fluxomics study requires measuring amino acids for M, M+1, M+2, and M+3 prioritizing glycine, serine, aspartate, asparagine, ornithine, citrulline, then as many as possible using a diamond hydride column and LCMS method (see CEvans).
Institute
University of Michigan
DepartmentBiomedical Research Core Facilities
LaboratoryMetabolomics core
Last NameKachman
First NameMaureen
Address6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Email mkachman@umich.edu
Submit Date2015-01-21
Num Groups9
Total Subjects27
Raw Data AvailableYes
Raw Data File Type(s).xml,.xsd,.bin, .cd,.cG
Uploaded File Size3.3 G
Analysis Type DetailGC/LC-MS
Release Date2015-04-24
Release Version1
Maureen Kachman Maureen Kachman
https://dx.doi.org/10.21228/M85C7W
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Project:

Project ID:PR000129
Project DOI:doi: 10.21228/M85C7W
Project Title:Metabolic effects of Noxa
Project Summary:labeled glutamine metabolomics on Noxa expressing cells
Institute:University of Minnesota
Department:Masonic Cancer Center
Laboratory:Kelekar Lab
Last Name:Kelekar
First Name:Ameeta
Address:420 Delaware Street SE, Minneapolis, MN 55455
Email:ameeta@umn.edu
Phone:612-626-2358

Subject:

Subject ID:SU000173
Subject Type:Human
Subject Species:Homo sapiens
Taxonomy ID:9606
Species Group:Human

Factors:

Subject type: Human; Subject species: Homo sapiens (Factor headings shown in green)

mb_sample_id local_sample_id Type Treatment
SA008406S00017122Noxa high clone 13C labeled glucose
SA008407S00017124Noxa high clone 13C labeled glucose
SA008408S00017123Noxa high clone 13C labeled glucose
SA008409S00017133Noxa high clone 15N alpha nitrogen labeled glutamine
SA008410S00017131Noxa high clone 15N alpha nitrogen labeled glutamine
SA008411S00017132Noxa high clone 15N alpha nitrogen labeled glutamine
SA008412S00017140Noxa high clone 15N amide nitrogen labeled glutamine
SA008413S00017142Noxa high clone 15N amide nitrogen labeled glutamine
SA008414S00017141Noxa high clone 15N amide nitrogen labeled glutamine
SA008415S00017130Noxa high plasmid 13C labeled glucose
SA008416S00017129Noxa high plasmid 13C labeled glucose
SA008417S00017128Noxa high plasmid 13C labeled glucose
SA008418S00017138Noxa high plasmid 15N alpha nitrogen labeled glutamine
SA008419S00017139Noxa high plasmid 15N alpha nitrogen labeled glutamine
SA008420S00017137Noxa high plasmid 15N alpha nitrogen labeled glutamine
SA008421S00017147Noxa high plasmid 15N amide nitrogen labeled glutamine
SA008422S00017146Noxa high plasmid 15N amide nitrogen labeled glutamine
SA008423S00017148Noxa high plasmid 15N amide nitrogen labeled glutamine
SA008424S00017125Noxa low clone 13C labeled glucose
SA008425S00017127Noxa low clone 13C labeled glucose
SA008426S00017126Noxa low clone 13C labeled glucose
SA008427S00017135Noxa low clone 15N alpha nitrogen labeled glutamine
SA008428S00017134Noxa low clone 15N alpha nitrogen labeled glutamine
SA008429S00017136Noxa low clone 15N alpha nitrogen labeled glutamine
SA008430S00017143Noxa low clone 15N amide nitrogen labeled glutamine
SA008431S00017145Noxa low clone 15N amide nitrogen labeled glutamine
SA008432S00017144Noxa low clone 15N amide nitrogen labeled glutamine
Showing results 1 to 27 of 27

Collection:

Collection ID:CO000159
Sample Type:Cell Extracts

Treatment:

Treatment ID:TR000179

Sample Preparation:

Sampleprep ID:SP000173
Sampleprep Protocol Filename:Fluxomics_analysis_protocol-2015-03-11.docx

Combined analysis:

Analysis ID AN000245 AN000246
Analysis type MS MS
Chromatography type HILIC GC
Chromatography system Agilent 1260 Agilent 7890A
Column Phenomenex Luna NH2 (150 x 1mm, 3um) Agilent DB5-MS (30m × 0.25mm, 0.25um)
MS Type ESI EI
MS instrument type QTOF Single quadrupole
MS instrument name Agilent 6520 QTOF Agilent 5975C
Ion Mode NEGATIVE POSITIVE
Units Area Area

Chromatography:

Chromatography ID:CH000170
Methods ID:AQM010
Methods Filename:QTOF-002-HILIC-35min-1mm-No_Insert.m
Instrument Name:Agilent 1260
Column Name:Phenomenex Luna NH2 (150 x 1mm, 3um)
Chromatography Type:HILIC
  
Chromatography ID:CH001302
Methods ID:AQM011
Methods Filename:ALPHA_KETO_ACIDS-FULL.M.zip
Instrument Name:Agilent 7890A
Column Name:Agilent DB5-MS (30m × 0.25mm, 0.25um)
Chromatography Type:GC

MS:

MS ID:MS000195
Analysis ID:AN000245
Instrument Name:Agilent 6520 QTOF
Instrument Type:QTOF
MS Type:ESI
Ion Mode:NEGATIVE
  
MS ID:MS000196
Analysis ID:AN000246
Instrument Name:Agilent 5975C
Instrument Type:Single quadrupole
MS Type:EI
Ion Mode:POSITIVE
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