PXD022511 is an
original dataset announced via ProteomeXchange.
Dataset Summary
Title | Reimagining high-throughput profiling of reactive cysteines for cell-based screening of large electrophile libraries |
Description | Current methods for measuring amino-acid side-chain reactivity lack the throughput needed to screen large chemical libraries for interactions across the proteome. Here we redesigned the workflow for activity-based protein profiling of reactive cysteine residues by using a smaller desthiobiotin-based probe, sample multiplexing, reduced protein starting amounts, and software to boost data acquisition in real-time on the mass spectrometer. Our method, Streamlined Cysteine Activity-Based Protein Profiling (SLC-ABPP), achieved a 42-fold improvement in sample throughput, corresponding to profiling library members at a depth of >8,000 reactive cysteine sites in 18 min per compound. We applied it to identifying the proteome-wide targets of a covalent inhibitor of mutant KRASG12C and of ibrutinib, a covalent inhibitor of BTK. In addition, we created a resource of cysteine reactivity to 285 electrophiles in three human cell lines , which includes >20,000 cysteines in >6,000 proteins per cell line. The goal of proteome-wide profiling of cysteine reactivity across thousand-member libraries under several cellular contexts is now within reach. |
HostingRepository | PRIDE |
AnnounceDate | 2021-01-26 |
AnnouncementXML | Submission_2021-01-25_22:59:06.xml |
DigitalObjectIdentifier | |
ReviewLevel | Peer-reviewed dataset |
DatasetOrigin | Original dataset |
RepositorySupport | Unsupported dataset by repository |
PrimarySubmitter | Miljan Kuljanin |
SpeciesList | scientific name: Mus musculus (Mouse); NCBI TaxID: 10090; scientific name: Homo sapiens (Human); NCBI TaxID: 9606; |
ModificationList | TMT6plex-126 reporter+balance reagent acylated residue; iodoacetic acid derivatized residue |
Instrument | Orbitrap Fusion Lumos |
Dataset History
Revision | Datetime | Status | ChangeLog Entry |
0 | 2020-11-13 03:13:42 | ID requested | |
⏵ 1 | 2021-01-25 22:59:07 | announced | |
Publication List
Kuljanin M, Mitchell DC, Schweppe DK, Gikandi AS, Nusinow DP, Bulloch NJ, Vinogradova EV, Wilson DL, Kool ET, Mancias JD, Cravatt BF, Gygi SP, Reimagining high-throughput profiling of reactive cysteines for cell-based screening of large electrophile libraries. Nat Biotechnol, 39(5):630-641(2021) [pubmed] |
Keyword List
submitter keyword: chemical proteomics, reactive cysteines |
Contact List
Steven P. Gygi |
contact affiliation | Department of Cell Biology, Harvard Medical School, Boston MA, USA |
contact email | steven_gygi@hms.harvard.edu |
lab head | |
Miljan Kuljanin |
contact affiliation | Harvard Medical School |
contact email | miljan_kuljanin@hms.harvard.edu |
dataset submitter | |
Full Dataset Link List
Dataset FTP location
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PRIDE project URI |
Repository Record List
[ + ]
[ - ]
- PRIDE
- PXD022511
- Label: PRIDE project
- Name: Reimagining high-throughput profiling of reactive cysteines for cell-based screening of large electrophile libraries