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PXD034790

PXD034790 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleNative holdup (nHU) to measure binding affinities from cell extracts
DescriptionCharacterizing macromolecular interactions is essential for understanding cellular processes, yet nearly all methods used to detect protein interactions from cells are qualitative. Here, we introduce a native holdup (nHU) approach that requires minimal sample preparation, investigates binding mechanisms, and quantifies equilibrium binding constants of protein interactions from cell extracts. We use nHU coupled with western blotting (nHU-WB) and mass spectrometry (nHU-MS) to explore interactions of SNX27, a cargo adaptor of the retromer complex. We find good agreement between affinities measured using minimal binding fragments and those measured directly from cell extracts using nHU. This challenges the unwritten paradigm stating that biophysical parameters like binding constants cannot be accurately determined from cells or cellular extracts. We discuss the strengths and limitations of nHU and provide simple protocols that can be implemented in most laboratories.
HostingRepositoryPRIDE
AnnounceDate2023-11-14
AnnouncementXMLSubmission_2023-11-14_08:37:08.733.xml
DigitalObjectIdentifier
ReviewLevelPeer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportUnsupported dataset by repository
PrimarySubmitterLuc Negroni
SpeciesList scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationListNo PTMs are included in the dataset
InstrumentLTQ Orbitrap Elite
Dataset History
RevisionDatetimeStatusChangeLog Entry
02022-06-21 07:22:37ID requested
12023-03-10 20:53:56announced
22023-11-14 08:37:10announced2023-11-14: Updated project metadata.
Publication List
Zambo B, Morlet B, Negroni L, Trave G, Gogl G, Native holdup (nHU) to measure binding affinities from cell extracts. Sci Adv, 8(51):eade3828(2022) [pubmed]
Keyword List
submitter keyword: protein interaction, mass spectrometry,native holdup
Contact List
Gilles Travé
contact affiliationIGBMC,UMR7104, Illkirch
contact emailtravegi@igbmc.fr
lab head
Luc Negroni
contact affiliationCNRS
contact emailluc.negroni@igbmc.fr
dataset submitter
Full Dataset Link List
Dataset FTP location
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PRIDE project URI
Repository Record List
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