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
| Study ID | ST000154 |
| Study Title | Use of Aspartate Dehydrogenase by cancer cells |
| Study Summary | The 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 |
| Department | Biomedical Research Core Facilities |
| Laboratory | Metabolomics core |
| Last Name | Kachman |
| First Name | Maureen |
| Address | 6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714 |
| mkachman@umich.edu | |
| Submit Date | 2015-01-21 |
| Num Groups | 9 |
| Total Subjects | 27 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | d |
| Uploaded File Size | 3.3 G |
| Analysis Type Detail | GC-MS/LC-MS |
| Release Date | 2015-04-24 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
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 |
|---|---|---|---|
| SA008406 | S00017122 | Noxa high clone | 13C labeled glucose |
| SA008407 | S00017124 | Noxa high clone | 13C labeled glucose |
| SA008408 | S00017123 | Noxa high clone | 13C labeled glucose |
| SA008409 | S00017133 | Noxa high clone | 15N alpha nitrogen labeled glutamine |
| SA008410 | S00017131 | Noxa high clone | 15N alpha nitrogen labeled glutamine |
| SA008411 | S00017132 | Noxa high clone | 15N alpha nitrogen labeled glutamine |
| SA008412 | S00017140 | Noxa high clone | 15N amide nitrogen labeled glutamine |
| SA008413 | S00017142 | Noxa high clone | 15N amide nitrogen labeled glutamine |
| SA008414 | S00017141 | Noxa high clone | 15N amide nitrogen labeled glutamine |
| SA008415 | S00017130 | Noxa high plasmid | 13C labeled glucose |
| SA008416 | S00017129 | Noxa high plasmid | 13C labeled glucose |
| SA008417 | S00017128 | Noxa high plasmid | 13C labeled glucose |
| SA008418 | S00017138 | Noxa high plasmid | 15N alpha nitrogen labeled glutamine |
| SA008419 | S00017139 | Noxa high plasmid | 15N alpha nitrogen labeled glutamine |
| SA008420 | S00017137 | Noxa high plasmid | 15N alpha nitrogen labeled glutamine |
| SA008421 | S00017147 | Noxa high plasmid | 15N amide nitrogen labeled glutamine |
| SA008422 | S00017146 | Noxa high plasmid | 15N amide nitrogen labeled glutamine |
| SA008423 | S00017148 | Noxa high plasmid | 15N amide nitrogen labeled glutamine |
| SA008424 | S00017125 | Noxa low clone | 13C labeled glucose |
| SA008425 | S00017127 | Noxa low clone | 13C labeled glucose |
| SA008426 | S00017126 | Noxa low clone | 13C labeled glucose |
| SA008427 | S00017135 | Noxa low clone | 15N alpha nitrogen labeled glutamine |
| SA008428 | S00017134 | Noxa low clone | 15N alpha nitrogen labeled glutamine |
| SA008429 | S00017136 | Noxa low clone | 15N alpha nitrogen labeled glutamine |
| SA008430 | S00017143 | Noxa low clone | 15N amide nitrogen labeled glutamine |
| SA008431 | S00017145 | Noxa low clone | 15N amide nitrogen labeled glutamine |
| SA008432 | S00017144 | Noxa low clone | 15N amide nitrogen labeled glutamine |
| Showing results 1 to 27 of 27 |
Collection:
| Collection ID: | CO000159 |
| Collection Summary: | - |
| Sample Type: | Cell Extracts |
Treatment:
| Treatment ID: | TR000179 |
Sample Preparation:
| Sampleprep ID: | SP000173 |
| Sampleprep Summary: | - |
| 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 |
| Acquisition Parameters File: | QTOF-002-HILIC-35min-1mm.m |
| Processing Parameters File: | EX00383-MassHunterQuant-DataAnalysis-Method.m |
| MS ID: | MS000196 |
| Analysis ID: | AN000246 |
| Instrument Name: | Agilent 5975C |
| Instrument Type: | Single quadrupole |
| MS Type: | EI |
| Ion Mode: | POSITIVE |
| Acquisition Parameters File: | ALPHA_KETO_ACIDS-FULL.M |
