PXD016014 is an
original dataset announced via ProteomeXchange.
Dataset Summary
Title | Inhibition of Ubiquitin Thioester Transfer by Targeting a Helix-in-Groove Interaction between E1 and E2 |
Description | In eukaryotes, E1 initiates the ubiquitin cascade by adenylation and thioesterification of the ubiquitin C-terminus and subsequent transfer of ubiquitin to E2 enzymes. A clinical-grade small molecule that binds to the E1 ATP binding site and covalently derivatizes the ubiquitin C-terminus effectively shuts down E1 enzymatic activity. However, mutation at or near the ATP binding site of E1 causes resistance, mandating alternative approaches to blocking what is otherwise a promising cancer target. Here, we identified a helix-in-groove interaction between the N-terminal alpha-1 helix of E2 and a pocket within the ubiquitin fold domain of E1 as a druggable site of protein interaction. By generating and optimizing stapled alpha-helical peptides (SAHs) modeled after the E2 alpha-1 helix, we achieve site-specific engagement of E1, induce a consequential conformational change, and effectively block E1 enzymatic activity, resulting in a generalized disruption of E2 ubiquitin-charging that suppresses ubiquitination of cellular proteins. Thus, we provide a blueprint for an alternative E1-targeting strategy for the treatment of cancer. Hydrogen exchange mass spectrometry was used to characterize the predominant E1 enzyme in mammals (UBE1, a 118 kDa multi-domain enzyme that catalyzes both ubiquitin adenylation and thioesterification) in the unbound state. We then interrogated the structural impact of UBE1 interaction with the stapled peptide SAH-UBE2A and several mutants. The observed peptide-induced exposure of the ubiquitin-fold domain (UFD) linker hinge in UBE1 was consistent with an inhibitory mechanism whereby SAH-UBE2A locks UBE1 into its proximal UFD conformation. |
HostingRepository | PRIDE |
AnnounceDate | 2020-08-18 |
AnnouncementXML | Submission_2020-08-22_16:45:25.xml |
DigitalObjectIdentifier | |
ReviewLevel | Peer-reviewed dataset |
DatasetOrigin | Original dataset |
RepositorySupport | Unsupported dataset by repository |
PrimarySubmitter | John R. Engen |
SpeciesList | scientific name: Homo sapiens (Human); NCBI TaxID: 9606; |
ModificationList | No PTMs are included in the dataset |
Instrument | Synapt MS |
Dataset History
Revision | Datetime | Status | ChangeLog Entry |
0 | 2019-10-25 02:34:19 | ID requested | |
1 | 2020-08-17 22:35:11 | announced | |
⏵ 2 | 2020-08-22 16:45:26 | announced | 2020-08-23: Updated publication reference for PubMed record(s): 32807965. |
Publication List
Keyword List
submitter keyword: Protein degradation, ubiquitin ligase, conformational change, hydrogen deuterium exchange, HDXMS |
Contact List
John R. Engen |
contact affiliation | Department of Chemistry & Chemical biology, Northeastern University |
contact email | j.engen@northeastern.edu |
lab head | |
John R. Engen |
contact affiliation | Northeastern University |
contact email | j.engen@northeastern.edu |
dataset submitter | |
Full Dataset Link List
Dataset FTP location
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PRIDE project URI |
Repository Record List
[ + ]
[ - ]
- PRIDE
- PXD016014
- Label: PRIDE project
- Name: Inhibition of Ubiquitin Thioester Transfer by Targeting a Helix-in-Groove Interaction between E1 and E2