PXD043402

PXD043402 is an original dataset announced via ProteomeXchange.

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

Revision	Datetime	Status	ChangeLog Entry
0	2023-06-29 10:05:28	ID requested
⏵ 1	2024-01-12 06:04:14	announced

10.1021/JACS.3C11780;

submitter keyword: Protein modifications, CuAAC, mass spectrometry, chemical proteomics, cysteine reactivity

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

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2024/01/PXD043402

PRIDE project URI

• PRIDE
  1. PXD043402
     1. Label: PRIDE project
     2. Name: Cu-Catalyzed Azide-Alkyne-Thiol Reaction Forms Ubiquitous Background in Chemical Proteomics Studies