Summary of Study ST002150
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 PR001363. The data can be accessed directly via it's Project DOI: 10.21228/M8J708 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.
Study ID | ST002150 |
Study Title | Sphingomyelin depletion inhibits CXCR4 dynamics and CXCL12-mediated directed cell migration in human T cells |
Study Summary | Sphingolipids, ceramides and cholesterol are integral components of cellular membranes, and they also play important roles in signal transduction by regulating the dynamics of membrane receptors through their effects on membrane fluidity. Here, we combined biochemical and functional assays with single-molecule dynamic approaches to demonstrate that the local lipid environment regulates CXCR4 organization and function and modulates chemokine-triggered directed cell migration. Prolonged treatment of T cells with neutral sphingomyelinase promoted the complete and sustained breakdown of sphingomyelins and the accumulation of the corresponding ceramides, which altered both membrane fluidity and CXCR4 nanoclustering and dynamics. Under these conditions CXCR4 retained some CXCL12-mediated signaling activity but failed to promote efficient directed cell migration. Our data underscore a critical role for the local lipid composition at the cell membrane in regulating the lateral mobility of chemokine receptors, and their ability to dynamically increase receptor density at the leading edge to promote efficient cell migration |
Institute | Universidad CEU San Pablo |
Last Name | Gonzalez-Riano |
First Name | Carolina |
Address | km 0, Universidad CEU-San Pablo Urbanización Montepríncipe. M-501 |
carolina.gonzalezriano@ceu.es | |
Phone | 646251045 |
Submit Date | 2022-04-22 |
Raw Data Available | Yes |
Raw Data File Type(s) | d |
Analysis Type Detail | LC-MS |
Release Date | 2022-05-09 |
Release Version | 1 |
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Sample Preparation:
Sampleprep ID: | SP002242 |
Sampleprep Summary: | For lipid extraction from Jurkat and T cell blasts, cell pellets were mixed with 200 µL of cold (-20°C) methanol:water (1:1, v/v) and sonicated with an ultrasonic homogenizer (UP200S, Hielscher Ultrasound Technology, HIELSCHER GmbH, Chamerau, Germany) for 16 bursts (0.5 second pulse) at 80% amplitude. Homogenates (100 µL) were mixed with 320 µL of cold (-20°C) methanol containing 1.6 ppm of sphinganine (d17:0) as the internal standard. Samples were then vortex-mixed for 2 minutes, followed by the addition of 80 µL of methyl tert-butyl ether. Subsequently, samples were vortex-mixed (1 hour, RT). After centrifugation (16,000 × g, 15°C, 10 minutes), samples were used for ultra-high performance liquid chromatography (UHPLC; Agilent 1290 Infinity II, Agilent Technologies Inc., Santa Clara, CA) coupled with (ESI) quadrupole time-of-flight (QTOF) mass spectrometry (MS) (Agilent 6546): 100 µL of each sample was divided between two UHPLC-MS vials with inserts (50 µL/each) for direct injection into the system for LC-MS analyses in positive and negative ionization modes. |