Summary of Study ST002412

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 PR001552. The data can be accessed directly via it's Project DOI: 10.21228/M8342Z 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.

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Study IDST002412
Study TitleMetabolic effects of the protein kinase R
Study TypeBiomedical research
Study SummarySpleen-derived macrophage from WT or Eif2ak2-/- (gene encoding PKR protein kinase) mice are treated with a synthetic RNA mimetic (polyinosinic:polycytidylic acid) to activate the kinase and metabolites were collected for analysis. The data identified 325 putative metabolites in the cell extracts, with a large number of significant differences between the Eif2ak2- /- and WT sample groups. Metabolite levels are predominantly suppressed in the WT compared to the Eif2ak2-/- cells, with depletion of specific metabolites in amino acid, carbohydrate, lipid and nucleotide pathways, while several amino acid metabolites were significantly elevated in the WT cells compared to the Eif2ak2-/-. The changes appear to delineate a pseudo-starvation response in the WT cells. Phosphate energy metabolism is altered with decreased creatine and phosphocreatine and a compensatory increase in phosphorylated guanidinoacetate in the WT compared to the Eif2ak2- /- cells. There appears to be a constraint in glycolysis in the WT cells, most clearly in the pentose phosphate pathway.
Institute
Hudson Institute of Medical Research
DepartmentCIIID
LaboratoryMolecular Immunology
Last NameSadler
First NameAnthony
Address27-31 Wright st, Clayton, VIC 3168
EmailAnthony.sadler@hudson.org.au
Phone+61 4 85722722
Submit Date2022-12-15
Num Groups2
Total SubjectsNA
Num MalesNA
Num FemalesNA
Study CommentsKO vs WT
PublicationsSuppression of the nucleic acid precursor ribose 5-phosphate by RNA-mediated antiviral immunity
Raw Data AvailableYes
Raw Data File Type(s)raw(Thermo)
Analysis Type DetailLC-MS
Release Date2023-01-04
Release Version1
Anthony Sadler Anthony Sadler
https://dx.doi.org/10.21228/M8342Z
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Sample Preparation:

Sampleprep ID:SP002507
Sampleprep Summary:Adherent macrophages were lifted with trypsin, suspended in PBS and 2x10 6 cells were collected by centrifugation. The cell pellet was rinsed in 10 mL ice cold 0.9% NaCl in water at 4 °C pelleted, then resuspended in 200 L of 4 °C extraction solvent (a 1:3:1 ratio of chloroform, methanol and water). The sample was subjected to three freeze-thaw cycles then mixed by vortex for 30 minutes at 4 °C and cell debris was removed by centrifugation at 20,000g for 10 minutes at 4 °C. The cleared supernatant was transferred (180 uL) and frozen at -80°C until LCMS analysis.
Extract Storage:-80℃
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