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PXD036953-1

PXD036953 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleThe interactome of flavin adenine dinucleotide (FAD) with RNA-binding proteins
DescriptionWe explored the interactome of flavin adenine dinucleotide (FAD) and the matrix of UV-immobilised FAD bound more than 3,000 proteins from SW-620 lysate. GO enrichment analysis showed that about 600 RNA-binding proteins were bound to FAD beads and, surprisingly, more than 40 proteins showed clear dose-dependent binding competition with nanomolar affinities indicating that their direct or indirect interaction with free FAD is both specific and strong. Among the few known consensus sequences for RNA-binding proteins, we selected the PUF motif 5'-UGUANAUA-3' to investigate whether FAD is binding the Pumilio homologs PUM1 and PUM2 in their RNA-binding pocket, as both these proteins showed dose-response behaviour in the FAD versus FAD-beads. Immobilisation of an oligomer containing the consensus sequence on NHS-activated Sepharose beads yielded PUM-beads that did enrich the Pumilio homologs. Oligomer vs FAD-beads and FAD vs PUM-beads together with the FAD vs FAD-beads dose-response competition indicated that the binding of FAD does not seem to influence the binding of the RNA. It is therefore most likely that FAD binding is allosteric for PUMs.
HostingRepositoryPRIDE
AnnounceDate2022-10-06
AnnouncementXMLSubmission_2022-10-06_04:17:25.997.xml
DigitalObjectIdentifier
ReviewLevelPeer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportUnsupported dataset by repository
PrimarySubmitterPolina Prokofeva
SpeciesList scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationListphosphorylated residue; iodoacetamide derivatized residue
InstrumentOrbitrap Fusion Lumos
Dataset History
RevisionDatetimeStatusChangeLog Entry
02022-09-22 05:53:00ID requested
12022-10-06 04:17:26announced
Publication List
Dataset with its publication pending
Keyword List
submitter keyword: PUM-RNA, FAD, competition pulldown, photo-immobilisation
Contact List
Bernhard Kuster
contact affiliationChair of Proteomics and Bioanalytics, TUM School of Life Sciences, Technical University of Munich, 85354 Freising, Germany
contact emailkuster@tum.de
lab head
Polina Prokofeva
contact affiliationChair of Proteomics and Bioanalytics, Technical University of Munich, Emil-Erlenmeyer-Forum 5, 85354 Freising, Germany
contact emailpolina.prokofeva@tum.de
dataset submitter
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Dataset FTP location
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