PXD016014

PXD016014 is an original dataset announced via ProteomeXchange.

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

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.

Cathcart AM, Bird GH, Wales TE, Herce HD, Harvey EP, Hauseman ZJ, Newman CE, Adhikary U, Prew MS, Oo T, Lee S, Engen JR, Walensky LD, Targeting a helix-in-groove interaction between E1 and E2 blocks ubiquitin transfer. Nat Chem Biol, 16(11):1218-1226(2020) [pubmed]

submitter keyword: Protein degradation, ubiquitin ligase, conformational change, hydrogen deuterium exchange, HDXMS

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

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PRIDE project URI

• PRIDE
  1. PXD016014
     1. Label: PRIDE project
     2. Name: Inhibition of Ubiquitin Thioester Transfer by Targeting a Helix-in-Groove Interaction between E1 and E2