PXD017943 is an
original dataset announced via ProteomeXchange.
Dataset Summary
Title | Phosphorylation ratio determination in fresh frozen and formalin-fixed paraffin embedded tissue with targeted mass spectrometry |
Description | Formalin-fixed paraffin embedded (FFPE) tissues are routinely prepared and collected for diagnostics in pathology departments. Therefore, FFPE tissues are the most accessible research sources in pathology archives. In this study we investigated whether we can apply a targeted and quantitative parallel reaction monitoring (PRM) method for FFPE tissue samples in a sensitive and reproducible way. Normal brain and glioblastoma multiforme (GBM) tissues were used to demonstrate the feasibility of our approach. Two analytical methods (or workflows) were used: PRM measurement of a tryptic digest without phosphopeptide enrichment (Direct-PRM) and after Fe-NTA phosphopeptide enrichment (Fe-NTA-PRM). Applying these two methods, the phosphorylation ratio could be determined for selected four peptide pairs that originate from Neuroblast differentiation-associated protein (AHNAK S5448-p), Calcium/calmodulin-dependent protein kinase type II subunit delta (CAMK2D T337-p), Eukaryotic translation initiation factor 4B (EIF4B S93-p) and Epidermal growth factor receptor (EGFR S1166-p). In normal brain FFPE tissues, using the Fe-NTA-PRM method, we were able to quantify the targeted phosphorylated peptides with a high degree of reproducibility (CV = 14%). Our results indicate that formalin fixation does not impede relative quantification of a phosphosite and its phosphorylation ratio in FFPE tissues. The developed methods open ways to study archival FFPE tissues. |
HostingRepository | PRIDE |
AnnounceDate | 2020-08-25 |
AnnouncementXML | Submission_2020-08-25_06:36:23.xml |
DigitalObjectIdentifier | https://dx.doi.org/10.6019/PXD017943 |
ReviewLevel | Peer-reviewed dataset |
DatasetOrigin | Original dataset |
RepositorySupport | Supported dataset by repository |
PrimarySubmitter | Lona Zeneyedpour |
SpeciesList | scientific name: Homo sapiens (Human); NCBI TaxID: 9606; |
ModificationList | Phospho; phosphorylated residue; Oxidation; Carbamidomethyl |
Instrument | Orbitrap Fusion Lumos |
Dataset History
Revision | Datetime | Status | ChangeLog Entry |
0 | 2020-03-09 04:45:02 | ID requested | |
⏵ 1 | 2020-08-25 06:36:27 | announced | |
Publication List
Zeneyedpour L, Stingl C, Dekker LJM, Mustafa DAM, Kros JM, Luider TM, Phosphorylation Ratio Determination in Fresh-Frozen and Formalin-Fixed Paraffin-Embedded Tissue with Targeted Mass Spectrometry. J Proteome Res, 19(10):4179-4190(2020) [pubmed] |
Keyword List
submitter keyword: Phosphoproteomics, PRM, Targeted mass spectrometry, FFPE, Phosphorylation ratio, Quantitative proteomics, Clinical proteomics, Cancer proteomics |
Contact List
Theo Luider |
contact affiliation | Head of Laboratory of Neuro-Oncology, Erasmus MC, Rotterdam |
contact email | t.luider@erasmusmc.nl |
lab head | |
Lona Zeneyedpour |
contact affiliation | Erasmus medical center |
contact email | l.zeneyedpour@erasmusmc.nl |
dataset submitter | |
Full Dataset Link List
Dataset FTP location
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PRIDE project URI |
Repository Record List
[ + ]
[ - ]
- PRIDE
- PXD017943
- Label: PRIDE project
- Name: Phosphorylation ratio determination in fresh frozen and formalin-fixed paraffin embedded tissue with targeted mass spectrometry