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PXD001609

PXD001609 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleValidation of a novel shotgun proteomic workflow for the discovery of protein-protein interactions: focus on ZNF521
DescriptionThe study of protein-protein interactions is increasingly relying on mass spectrometry (MS). The classical approach of separating immunoprecipitated proteins by SDS-PAGE followed by in-gel digestion is long and labour-intensive. Besides, it is difficult to integrate it with most quantitative MS-based workflows, except for stable isotopic labelling of amino acids in cell culture (SILAC). This work describes a fast, flexible and quantitative workflow for the discovery of novel proteinprotein interactions. A cleavable cross-linker, dithiobis[succinimidyl propionate] (DSP), is utilized to stabilize protein complexes before immunoprecipitation. Protein complex detachment from the antibody is achieved by limited proteolysis. Finally, protein quantitation is performed via 18O labelling. The workflow has been optimized concerning: (i) DSP concentration and (ii) incubation times for limited proteolysis, using the stem cell-associated transcription co-factor ZNF521 as a model target. The interaction of ZNF521 with the core components of the nuclear remodelling and histone deacetylase (NuRD) complex, already reported in the literature, was confirmed. Additionally, interactions with newly discovered molecular partners of potentially relevant functional role, such as ZNF423, Spt16, Spt5, were discovered and validated by Western blotting.
HostingRepositoryPRIDE
AnnounceDate2015-04-07
AnnouncementXMLSubmission_2015-04-07_10:42:36.xml
DigitalObjectIdentifierhttps://dx.doi.org/10.6019/PXD001609
ReviewLevelPeer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportSupported dataset by repository
PrimarySubmitterSHIBU KRISHNAN
SpeciesList scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationListNo PTMs are included in the dataset
InstrumentQ Exactive
Dataset History
RevisionDatetimeStatusChangeLog Entry
02014-12-15 03:00:46ID requested
12015-04-07 10:42:37announced
Publication List
Bernaudo F, Monteleone F, Mesuraca M, Krishnan S, Chiarella E, Scicchitano S, Cuda G, Morrone G, Bond HM, Gaspari M, Validation of a novel shotgun proteomic workflow for the discovery of protein-protein interactions: focus on ZNF521. J Proteome Res, 14(4):1888-99(2015) [pubmed]
Keyword List
curator keyword: Biological, Technical
submitter keyword: AP-MS
interactomics
18O labelling
ZNF521
ZNF423
Spt16
Spt5
NuRD core complex
proteomics
mass spectrometry
shotgun proteomics
Contact List
Professor Giovanni Cuda
contact affiliationUniversity Magna Graecia of Catanzaro
contact emailcuda@unicz.it
lab head
SHIBU KRISHNAN
contact affiliationUniversity Magna Graecia of Catanzaro
contact emailshibu@unicz.it
dataset submitter
Full Dataset Link List
Dataset FTP location
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PRIDE project URI
Repository Record List
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