PXD012631-2

PXD012631 is an original dataset announced via ProteomeXchange.

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	2024-10-22
AnnouncementXML	Submission_2024-10-22_05:07:14.593.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

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.

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]

submitter keyword: stress granule,Yeast, heatshock

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

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2020/06/PXD012631

PRIDE project URI

• PRIDE
  1. PXD012631
     1. Label: PRIDE project
     2. Name: Yeast Heatshock SILAC LC-MSMS