PXD008026

PXD008026 is an original dataset announced via ProteomeXchange.

Title	MudPIT analyses of whole cell extracts from Y1 murine adrenocortical carcinoma cell line stimulated with serum or FGF2
Description	Fibroblast growth factor 2 (FGF2) is well known as a promoter of cell proliferation, however, in some cellular contexts, it can induce cell cycle arrest or cell death. In Y1 murine adrenocortical carcinoma cell line, FGF2 induces G0/G1 transition but delays S-phase progression and permanently block cells in G2/M. To gain insights into the molecular mechanisms of this antiproliferative effect, we investigated the early proteomics alterations induced by this growth factor. We compared mass spectrometry-based quantitative proteomics analyses of Y1 cells stimulated, after 3 and 5 hours, with fetal bovine serum (FBS) or FGF2. Total protein extracts from Y1 cells stimulated with FBS or FBS plus FGF2 (10 ng/ml) for 0 h, 3 h, and 5 h were obtained in 3 biological replicates. One hundred and fifty micrograms of proteins were TCA precipitated and dissolved in 100 mM Tris-HCL pH 8.5, 8 M urea. The sample was reduced with 5 mM TCEP, incubated at room temperature for 30 min, and carboxymethylated by the addition of 10 mM of iodoacetamide. Proteins were digested with endoproteinase Lys-C for 4 h and then digested with Trypsin overnight at 37C. The peptides were loaded onto a 100 um three-phase capillary column packed with 8 cm of 5 um C18 reverse phase (Aqua; Phenomenex), followed by 3 cm of 5 um SCX resin (Partisphere SCX; Whatman) and an additional 1.5 cm of reverse phase. Columns loaded with the peptides were placed in-line with an Agilent 1100 quaternary HPLC connected with a LTQ-XL (Thermo) mass spectrometer equipped with a nanospray ionization source. Twelve automatic steps were used in the MudPIT run with increasing concentrations of ammonium acetate. Each full MS scan (400 m/z to 1600 m/z) was followed by 5 MS/MS events fragmented by CID.
HostingRepository	MassIVE
AnnounceDate	2018-07-16
AnnouncementXML	Submission_2018-07-16_12:43:55.xml
DigitalObjectIdentifier
ReviewLevel	Non peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Laurence Florens
SpeciesList	scientific name: Mus musculus; common name: house mouse; NCBI TaxID: 10090;
ModificationList	L-methionine sulfoxide; S-carboxamidomethyl-L-cysteine
Instrument	LTQ XL

Revision	Datetime	Status	ChangeLog Entry
0	2017-10-23 08:05:09	ID requested
⏵ 1	2018-07-16 12:43:55	announced

no publication

submitter keyword: Fibroblast growth factor 2, fosB, junB, src, replication, mice tumor cell

Laurence Florens
contact affiliation	The Stowers Institute for Medical Research
contact email	laf@stowers.org
lab head
Laurence Florens
contact affiliation	Stowers Institute for Medical Research
contact email	laf@stowers.org
dataset submitter

MassIVE dataset URI

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