PXD036953

PXD036953 is an original dataset announced via ProteomeXchange.

Title	The interactome of flavin adenine dinucleotide (FAD) with RNA-binding proteins
Description	We 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.
HostingRepository	PRIDE
AnnounceDate	2022-10-06
AnnouncementXML	Submission_2022-10-06_04:17:25.997.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Polina Prokofeva
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	phosphorylated residue; iodoacetamide derivatized residue
Instrument	Orbitrap Fusion Lumos

Revision	Datetime	Status	ChangeLog Entry
0	2022-09-22 05:53:00	ID requested
⏵ 1	2022-10-06 04:17:26	announced

Dataset with its publication pending

submitter keyword: PUM-RNA, FAD, competition pulldown, photo-immobilisation

Bernhard Kuster
contact affiliation	Chair of Proteomics and Bioanalytics, TUM School of Life Sciences, Technical University of Munich, 85354 Freising, Germany
contact email	kuster@tum.de
lab head
Polina Prokofeva
contact affiliation	Chair of Proteomics and Bioanalytics, Technical University of Munich, Emil-Erlenmeyer-Forum 5, 85354 Freising, Germany
contact email	polina.prokofeva@tum.de
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD036953
     1. Label: PRIDE project
     2. Name: The interactome of flavin adenine dinucleotide (FAD) with RNA-binding proteins