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PXD014066

PXD014066 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleEvaluation and Optimization of Chemically-Cleavable Linkers for Quantitative Mapping of Small Molecule-Protein Interactomes
DescriptionNumerous reagents have been developed to enable chemical proteomic analysis of small molecule-protein interactomes. However, the performance of these reagents has not been systematically evaluated and compared. Herein, we report our efforts to conduct a parallel assessment of two widely-used chemically-cleavable linkers equipped with dialkoxydiphenylsilane (DADPS linker) and azobenzene (AZO linker) moieties. Profiling a cellular cysteinome using iodoacetamide alkyne probe demonstrated a significant discrepancy between the experimental results obtained through the application of each of the reagents. To better understand the source of observed discrepancy, a mass tolerant database search strategy using MSFragger software was performed. This resulted in identifying a previously unreported artifactual modification on the residual mass of the azobenzene linker. Furthermore, we conducted a comparative analysis of enrichment modes using both cleavable linkers. This effort determined that enrichment of proteolytic digests yielded a far greater number of identified cysteine residues than the enrichment conducted prior to protein digest. Inspired by recent studies where multiplexed quantitative labeling strategies were applied to cleavable biotin linkers, we combined this further optimized protocol using the DADPS cleavable linker with tandem mass tag (TMT) labeling to profile the FDA-approved covalent EGFR kinase inhibitor dacomitinib against the cysteinome of an epidermoid cancer cell line. Our analysis resulted in the detection and quantification of over 10,000 unique cysteine residues, a nearly 3-fold increase over previous studies that used cleavable biotin linkers for enrichment. Critically, cysteine residues corresponding to proteins directly as well as indirectly modulated by dacomitinib treatment were identified. Overall, our study suggests that the dialkoxydiphenylsilane linker could be broadly applied wherever chemically cleavable linkers are required for chemical proteomic characterization of cellular proteomes.
HostingRepositoryPRIDE
AnnounceDate2024-10-22
AnnouncementXMLSubmission_2024-10-22_04:54:10.900.xml
DigitalObjectIdentifier
ReviewLevelPeer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportUnsupported dataset by repository
PrimarySubmitterAdam Rabalski
SpeciesList scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationListsulfated residue; monohydroxylated residue; iodoacetamide derivatized residue
InstrumentOrbitrap Fusion Lumos; Orbitrap Fusion
Dataset History
RevisionDatetimeStatusChangeLog Entry
02019-05-29 09:00:11ID requested
12019-08-20 23:31:41announced
22024-10-22 04:54:19announced2024-10-22: Updated project metadata.
Publication List
Rabalski AJ, Bogdan AR, Baranczak A, Evaluation of Chemically-Cleavable Linkers for Quantitative Mapping of Small Molecule-Cysteinome Reactivity. ACS Chem Biol, 14(9):1940-1950(2019) [pubmed]
10.1021/acschembio.9b00424;
Keyword List
submitter keyword: azobenzene, dacomitinib, TMT, cleavable biotin linker, dialkoxydiphenylsilane
Contact List
Aleksandra Baranczak
contact affiliationDiscovery Platform Technologies, AbbVie
contact emailaleksandra.baranczak@abbvie.com
lab head
Adam Rabalski
contact affiliationDiscovery Platform Technologies, AbbVie
contact emailadam.rabalski@abbvie.com
dataset submitter
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Dataset FTP location
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