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PXD034639

PXD034639 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleAn improved TurboID-based proximity labeling pipeline in C. elegans by biochemical depletion of endogenously biotinylated carboxylases
DescriptionTo review an improved proximity-labeling strategy that uses the improved E. coli biotin ligase TurboID to characterize C. elegans protein complexes, the interactome of a presynaptic active zone protein, ELKS-1, was analyzed as proof of principle. A significant constraint on the sensitivity of TurboID-based proximity labeling is the presence of abundant, endogenously biotinylated proteins that take up bandwidth in the mass spectrometer, notably carboxylases that use biotin as a co-factor. We developed ways to remove these carboxylases prior to streptavidin purification and mass spectrometry, by engineering their corresponding genes to add a C-terminal His10 tag. This allows us to deplete them from C. elegans lysates using immobilized metal affinity chromatography (IMAC).
HostingRepositoryPRIDE
AnnounceDate2023-11-14
AnnouncementXMLSubmission_2023-11-14_08:56:43.683.xml
DigitalObjectIdentifier
ReviewLevelPeer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportUnsupported dataset by repository
PrimarySubmitterWeiQiang Chen
SpeciesList scientific name: Caenorhabditis elegans; NCBI TaxID: 6239;
ModificationListbiotinylated residue; acetylated residue; monohydroxylated residue; iodoacetamide derivatized residue
InstrumentOrbitrap Exploris 480
Dataset History
RevisionDatetimeStatusChangeLog Entry
02022-06-19 04:46:00ID requested
12022-09-12 11:47:11announced
22023-11-14 08:56:44announced2023-11-14: Updated project metadata.
Publication List
Dataset with its publication pending
Keyword List
submitter keyword: synapse, IMAC, APEX2, proximity-dependent protein labeling, HRP, biotinylated carboxylase depletion,TurboID, Caenorhabditis elegans
Contact List
Mario de Bono
contact affiliationInstitute of Science and Technology Austria (ISTA), Am Campus 1, 3400 Klosterneuburg, Austria
contact emailmdebono@ista.ac.at
lab head
WeiQiang Chen
contact affiliationMass Spectrometry Facility, Max Perutz Laboratories Support GmbH
contact emailchenweiqiang@gmail.com
dataset submitter
Full Dataset Link List
Dataset FTP location
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PRIDE project URI
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