Summary of Study ST001674
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 PR001077. The data can be accessed directly via it's Project DOI: 10.21228/M8GT3N This work is supported by NIH grant, U2C- DK119886.
See: https://www.metabolomicsworkbench.org/about/howtocite.php
Study ID | ST001674 |
Study Title | Untargeted phospholipid analysis of HeLa silenced for or overexpressing GOLPH3 |
Study Summary | A group of sequentially-acting enzymes operating at the branchpoint among sphingolipid synthetic pathways binds the Golgi-localised oncoprotein GOLPH3. GOLPH3 sorts these enzymes into vesicles for intra-Golgi retro-transport, acting as a component of the cisternae inter-conversion mechanisms. Through these effects, GOLPH3 controls the sub-Golgi localisation, and the lysosomal degradation rate of specific enzymes. Here we evaluated the impact of overexpressing or silencing GOLPH3 on the glycerophospholipid composition of HeLa cells by untargeted lipid analysis. |
Institute | École polytechnique fédérale de Lausanne (EPFL) |
Department | IBI |
Laboratory | UPDANGELO |
Last Name | D'Angelo |
First Name | Giovanni |
Address | Station 15, Lausanne, Vaud, 1015, Switzerland |
giovanni.dangelo@epfl.ch | |
Phone | +41 216934276 |
Submit Date | 2021-01-29 |
Raw Data Available | Yes |
Raw Data File Type(s) | raw(Thermo) |
Analysis Type Detail | LC-MS |
Release Date | 2021-02-26 |
Release Version | 1 |
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Sample Preparation:
Sampleprep ID: | SP001757 |
Sampleprep Summary: | Total lipid extracts were prepared using a standard MTBE protocol followed by a methylamine treatment for total lipid analysis by mass spectrometry. Briefly, cell pellets or viral fractions were resuspended in 100 μL H2O. 360 μL methanol and 1.2 mL of MTBE were added and samples were placed for 10 min on a vortex at 4 C followed by incubation for 1 h at room temperature on a shaker. Phase separation was induced by addition of 200 μL of H2O. After 10 min at room temperature, samples were centrifuged at 1000 g for 10 min. The upper (organic) phase was transferred into a glass tube and the lower phase was re-extracted with 400 μL artificial upper phase [MTBE/methanol/H2O (10:3:1.5, v/v/v)]. The combined organic phases were dried in a vacuum concentrator. Lipids where then resuspended in 500 μL of ChCl3. |