PXD043402 is an
original dataset announced via ProteomeXchange.
Dataset Summary
Title | Cu-Catalyzed Azide-Alkyne-Thiol Reaction Forms Ubiquitous Background in Chemical Proteomics Studies |
Description | We report here a Cu-catalyzed azide-alkyne-thiol reaction forming thiotriazoles as the major by-product under widely used bioorthogonal protein labelling “click” conditions. The development of Cu(I)-catalyzed azide-alkyne cycloaddition (CuAAC) had tremendous impact on many biological discoveries. However, the considered chemoselectivity of CuAAC is hampered by high reactivity of cysteine free thiols yielding thiotriazole protein conjugates. The reaction by-products generate false positive protein hits in “functional” proteomic studies. The reported detail investigation of conjugates between chemical probes containing terminal alkynes, azide-tags and cell lysates reveals formation of thiotriazoles, which can be readily detected by in-gel fluorescence scanning or after peptide and protein enrichment by MS-based proteomics. The produced fluorescent bands or enriched proteins may not result from the important enzymatically driven reaction and can be falsely assigned as hits. This study provides a complete list of the most common background proteins. The knowledge of this previously overlooked reactivity now leads to improved experimental design. Here, we present modified CuAAC conditions, which avoids the undesired product formation and diminishes the background. |
HostingRepository | PRIDE |
AnnounceDate | 2024-01-12 |
AnnouncementXML | Submission_2024-01-12_06:04:14.363.xml |
DigitalObjectIdentifier | |
ReviewLevel | Peer-reviewed dataset |
DatasetOrigin | Original dataset |
RepositorySupport | Unsupported dataset by repository |
PrimarySubmitter | Andreas Wiest |
SpeciesList | scientific name: Homo sapiens (Human); NCBI TaxID: 9606; |
ModificationList | iodoacetamide derivatized residue |
Instrument | Orbitrap Eclipse |
Dataset History
Revision | Datetime | Status | ChangeLog Entry |
0 | 2023-06-29 10:05:28 | ID requested | |
⏵ 1 | 2024-01-12 06:04:14 | announced | |
Publication List
Keyword List
submitter keyword: Protein modifications, CuAAC, mass spectrometry, chemical proteomics, cysteine reactivity |
Contact List
Pavel Kielkowski |
contact affiliation | Department of Chemistry, LMU Munich, Würmtalstr. 201, 81375, Munich, Germany |
contact email | pavel.kielkowski@cup.lmu.de |
lab head | |
Andreas Wiest |
contact affiliation | Department of Chemistry, LMU Munich, Würmtalstr. 201, 81375, Munich, Germany |
contact email | andreas.wiest@cup.lmu.de |
dataset submitter | |
Full Dataset Link List
Dataset FTP location
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PRIDE project URI |
Repository Record List
[ + ]
[ - ]
- PRIDE
- PXD043402
- Label: PRIDE project
- Name: Cu-Catalyzed Azide-Alkyne-Thiol Reaction Forms Ubiquitous Background in Chemical Proteomics Studies