PXD000488 is an
original dataset announced via ProteomeXchange.
Dataset Summary
Title | Comparative Proteomic profiling of the rostral band of LAL muscle from SMA mice and litter mate controls |
Description | Mouse sample preparation: 'Severe' SMA mice (Smn-/-;SMN2+/+) and wild-type (Smn+/+;SMN2+/+) littermates at postnatal day 5 (P5) were sacrificed by chilling on ice and decapitation. Levator auris longus (LAL, from the back of the neck) muscles were dissected in oxygenated mammalian physiological saline, as previously described [PMID: 19640925]. LAL muscles were separated into rostral and caudal bands and quickly frozen on dry ice. The rostral band of LAL from each mouse was stored at -80C until sufficient tissue was collected for proteomics analysis. Proteomic analysis: Protein was extracted in MEBC Buffer (50 mM TRIS, 100 mM NaCl, 5 mM NaEDTA, 5 mM NaEGTA, 40 mM beta-glycerophosphate, 100 mM sodium fluoride (NaF), 100 mM sodium orthovanadate, 0.25% NP-40, 1 Roche 'complete' protease inhibitor tablet, pH 7.4). Protein concentration was determined by BCA P5 WT and KO Rostral). Then 10ug aliquots of each muscle type assay according to manufacturers instructions on solubilised muscle (were reduced with 10 mM DTT and alkylated with 50 mM iodoacetamide prior to digestion with trypsin (Roche, sequencing grade) overnight at 30C. Technical replicates (3 x 2.5 ug) of each digested muscle type were injected onto a nLCMS/ MS system (Ultimate 3000 (Dionex) coupled to a LTQ Orbitrap XL (Thermo Scientific). The peptides from each digest were separated over a 65 min linear gradient from 5-35% acetonitrile in 0.1% formic acid. The LTQ Orbitrap XL was configured with a TOP 5 methodology comprising a 60K resolution FT-MS full scan followed by IT-MS/MS scans for the 5 most intense peptide ions. The raw data was then imported into Progenesis LCMS for label free differential analysis and subsequent identification and quantification of relative ion abundance ratios, both up-regulated and down-regulated. Following alignment of MS data, principal component analysis and preliminary filtering (power >80%,P > 0.05), data was exported from Progenesis as a single mgf file per time point. These files were then used to identify individual peptide sequences using the Swiss-Prot database via Mascot Daemon (V2.4.0) due to the large file size. As an indication of identification certainty, the false discovery rate for peptide matches above identity threshold was 3.34% for P5. |
HostingRepository | PRIDE |
AnnounceDate | 2014-07-28 |
AnnouncementXML | Submission_2014-07-28_00:49:26.xml |
DigitalObjectIdentifier | https://dx.doi.org/10.6019/PXD000488 |
ReviewLevel | Peer-reviewed dataset |
DatasetOrigin | Original dataset |
RepositorySupport | Supported dataset by repository |
PrimarySubmitter | Douglas Lamont |
SpeciesList | scientific name: Mus musculus (Mouse); NCBI TaxID: 10090; |
ModificationList | monohydroxylated residue; acetylated residue; iodoacetamide derivatized residue; deaminated residue; dihydroxylated residue |
Instrument | instrument model: LTQ Orbitrap XL; LTQ Orbitrap XL |
Dataset History
Revision | Datetime | Status | ChangeLog Entry |
0 | 2013-10-02 06:45:27 | ID requested | |
1 | 2013-10-09 03:38:54 | announced | |
1 | 2013-12-19 04:33:04 | announced | |
⏵ 2 | 2014-07-28 00:49:26 | announced | Updated project metadata. |
Publication List
Dataset with its publication pending |
Keyword List
submitter keyword: Mouse LAL muscle SMA LTQ Orbitrap XL |
Contact List
Douglas Lamont |
contact affiliation | School of Life Sciences |
contact email | d.j.lamont@dundee.ac.uk |
dataset submitter | |
Full Dataset Link List
Dataset FTP location
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PRIDE project URI |
Repository Record List
[ + ]
[ - ]
- PRIDE
- PXD000488
- Label: PRIDE project
- Name: Comparative Proteomic profiling of the rostral band of LAL muscle from SMA mice and litter mate controls