Summary of Study ST001786
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 PR001135. The data can be accessed directly via it's Project DOI: 10.21228/M80D82 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 | ST001786 |
| Study Title | Multi-omics analysis of glucose-mediated signaling by a moonlighting Gβ protein Asc1/RACK1 |
| Study Type | Untargeted UPLC-MS Metabolomics Analysis |
| Study Summary | While much is known about glucose metabolism in yeast, less is known about the receptors and signaling pathways that indicate glucose availability. We obtained metabolic profiles for wildtype and 16 mutants affecting the yeast glucose sensing pathway, comparing 0.05% glucose vs 10 min after glucose addition to 2%. First, we determined that the G protein-coupled receptor (Gpr1/Gpa2) directs early events in glucose utilization while the transceptors (Snf3/Rgt2) regulate subsequent processes and downstream products of glucose metabolism. Whereas the large G protein transmits the signal from its cognate receptor, Ras2 (but not Ras1) integrates responses from both receptor pathways. Second, we determined the relative contributions of the G protein α (Gpa2) and β (Asc1) subunits to glucose-initiated processes. We determined that Gpa2 is primarily involved in regulating carbohydrate metabolism while Asc1 is primarily involved in amino acid metabolism. Both proteins are involved in regulating purine metabolism. Collectively, our analysis reveals the molecular basis for glucose detection and the earliest events of glucose-dependent signal transduction in yeast. |
| Institute | University of North Carolina at Chapel Hill |
| Department | Nutrition |
| Laboratory | Metabolomics and Exposome Laboratory, Nutrition Research Institute, UNC Chapel Hill |
| Last Name | Sumner |
| First Name | Susan |
| Address | 500 Laureate Way, Nutrition Research Institute, UNC Chapel Hill |
| susan_sumner@unc.edu | |
| Phone | (919) 622-4456 |
| Submit Date | 2021-05-10 |
| Num Groups | 28 |
| Total Subjects | 192 |
| Raw Data Available | Yes |
| Raw Data File Type(s) | raw(Thermo) |
| Analysis Type Detail | LC-MS |
| Release Date | 2021-06-11 |
| Release Version | 1 |
Select appropriate tab below to view additional metadata details:
Sample Preparation:
| Sampleprep ID: | SP001869 |
| Sampleprep Summary: | Frozen cell pellets were resuspended with extraction reagent (8:2 methanol-water solution) to 3x108 cell/mL and then transferred into 2 mL ceramic bead MagNalyser tubes. Blank samples were prepared by adding 1300 μL of extraction reagent with no cells to a MagNalyser tube with ceramic beads. Tubes were subjected to homogenization, with Bead Ruptor Elite Bead Mill Homogenizer (OMNI International) at 6.0 m/s for 40 sec in 2 cycles at room temperature. This step was repeated twice. All samples were then centrifuged at 16,000 xg for 10 min at 4 °C. 500 μL of the supernatant was transferred into low-bind 1.7 mL microfuge tubes. Total pools were made by combining an additional 65 μL of the supernatant from each sample and then aliquoting this mixture into low-bind 1.7 mL tubes at a volume of 500 μL. The remaining supernatant was stored at -80 °C for repeat experiments if necessary. For all experimental samples, pooled samples and blanks were dried using a speedvac vacuum concentrator overnight. Dried samples were stored at -80 °C. Before LC-MS analysis, 100 μL of reconstitution buffer (95:5 water:methanol with 500 ng/mL tryptophan d-5) was added to each dried sample. All tubes were vortexed at 5000 rpm for 10 min and then centrifuged at room temperature at 16,000 xg for 4 min. Supernatant was transferred into autosampler vials for LC-MS. |
| Processing Storage Conditions: | 4℃ |
| Extract Storage: | -80℃ |
