PXD012631 is an
original dataset announced via ProteomeXchange.
Dataset Summary
Title | Yeast Heatshock SILAC LC-MSMS . |
Description | Yeast cells were incubated at 25˚C, unless otherwise stated. For SILAC, cells were grown in light labeled (0.03mg/ml Lys0, 0.02mg/ml Arg0) or heavy labeled (0.03mg/ml Lys4, 0.02mg/ml Arg6) amino acid in synthetic defined medium with 2% dextrose for at least 7 generations. heat shock was performed in a shaking water bath at 45˚C for 20 minutes before harvest. Equal OD600 of cells from both labels were collected at mid log phase (OD¬600=0.8-1) by centrifugation at 3,220xg for 5 minutes at 4˚C, washed twice with cold 1×TBS (50mM Tris pH 7.5, 150mM NaCl), mixed, and re-suspended in equal volume of 2×Native lysis buffer (200mM Tris pH 7.5, 150mM NaCl, 2mM PMSF, 2×Protease Inhibitor Cocktails (Sigma-Aldrich), 2mM 1,10-Phenanthroline). Re-suspended cells were snap-frozen drop by drop in liquid nitrogen. The resulting cells pellets were lysed by cyro-grinding in liquid nitrogen with a mortar and pestle mounted on an electric driver (OPS Diagnostics). The lysate was thawed on ice and further diluted 3 times by adding ice-cold 1×Native lysis buffer and 1% NP40 (final concentration). Total cell lysate was pre-cleared twice by centrifugation at 1,000×g at 4˚C for 15min. Pellet fraction was separated by centrifugation at 16,100×g at 4˚C for 15min. The insoluble protein pellet was washed twice with ice-cold 1×Native lysis buffer containing 1% NP40 and re-solubilized in 1×Laemmli Sample Buffer (62.5mM Tris-HCl pH6.8, 2% SDS, 10% Glycerol). Protein concentrations were determined by using BioRad DC™ Protein Assay. |
HostingRepository | PRIDE |
AnnounceDate | 2024-10-07 |
AnnouncementXML | Submission_2024-10-07_10:23:38.876.xml |
DigitalObjectIdentifier | |
ReviewLevel | Peer-reviewed dataset |
DatasetOrigin | Original dataset |
RepositorySupport | Unsupported dataset by repository |
PrimarySubmitter | Mang Zhu |
SpeciesList | scientific name: Saccharomyces cerevisiae (Baker's yeast); NCBI TaxID: 4932; |
ModificationList | iodoacetamide derivatized residue |
Instrument | impact II |
Dataset History
Revision | Datetime | Status | ChangeLog Entry |
0 | 2019-02-08 02:32:55 | ID requested | |
⏵ 1 | 2024-10-07 10:23:39 | announced | |
2 | 2024-10-22 05:07:21 | announced | 2024-10-22: Updated project metadata. |
Publication List
10.1242/jcs.244657; |
Zhu M, Kuechler ER, Zhang J, Matalon O, Dubreuil B, Hofmann A, Loewen C, Levy ED, Gsponer J, Mayor T, . J Cell Sci, 133(13):(2020) [pubmed] |
Keyword List
submitter keyword: stress granule,Yeast, heatshock |
Contact List
Thibault Mayor |
contact affiliation | Michael Smith Laboratories, Department of Biochemistry and Molecular Biology, University of British Columbia, Vancouver, BC, Canada |
contact email | mayor@msl.ubc.ca |
lab head | |
Mang Zhu |
contact affiliation | UBC |
contact email | mzhu@gsat.ubc.ca |
dataset submitter | |
Full Dataset Link List
Dataset FTP location
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PRIDE project URI |
Repository Record List
[ + ]
[ - ]
- PRIDE
- PXD012631
- Label: PRIDE project
- Name: Yeast Heatshock SILAC LC-MSMS