PXD006868

PXD006868 is an original dataset announced via ProteomeXchange.

Title	A linear diubiquitin-based probe for efficient and selective detection of the deubiquitinating enzyme OTULIN
Description	The methionine1 (M1)-specific deubiquitinase OTULIN acts as a negative regulator of NF-ΚB signaling and immune homeostasis. By replacing G76 in distal ubiquitin by dehydroalanine we designed the diubiquitin (diUb) probe UbG76Dha-Ub (OTULIN ABP) that couples to the catalytic site of OTULIN and thereby captures OTULIN in its active conformation. OTULIN ABP displays high selectivity for OTULIN and is not labeling to other M1-specific DUB including CYLD. In fact, labeling of USP5/IsopeptidaseT was the only detectable cross-reactivity. Further, ATP-dependent association of UbG76Dha-Ub probe with E1 activating enzymes induces assembly of polyOTULIN ABP chains, but USP5 labelling and E1 binding was abolished by deleting the C-terminal glycine in proximal ubiquitin (UbG76Dha-UbΔG76: OTULIN ABPΔG76). Pull-downs demonstrate that substrate-bound OTULIN associates with the linear ubiquitin chain assembly complex (LUBAC). Thus, we present a highly selective DUB for OTULIN that will facilitate studying the cellular function of this critical deubiquitinase.
HostingRepository	PRIDE
AnnounceDate	2017-09-19
AnnouncementXML	Submission_2017-09-19_04:48:06.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Philip Charles
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	monohydroxylated residue; iodoacetamide derivatized residue; deamidated residue
Instrument	Q Exactive HF

Revision	Datetime	Status	ChangeLog Entry
0	2017-07-05 07:39:14	ID requested
⏵ 1	2017-09-19 04:48:07	announced

Weber A, Elliott PR, Pinto-Fernandez A, Bonham S, Kessler BM, Komander D, El Oualid F, Krappmann D, A Linear Diubiquitin-Based Probe for Efficient and Selective Detection of the Deubiquitinating Enzyme OTULIN. Cell Chem Biol, 24(10):1299-1313.e7(2017) [pubmed]

curator keyword: Technical, Biological

submitter keyword: Human, OTULIN

Benedikt M Kessler
contact affiliation	Target Discovery Institute, Nuffield Department of Medicine, University of Oxford, Roosevelt Drive, Oxford OX3 7FZ, UK
contact email	benedikt.kessler@ndm.ox.ac.uk
lab head
Philip Charles
contact affiliation	University of Oxford
contact email	philip.charles@ndm.ox.ac.uk
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD006868
     1. Label: PRIDE project
     2. Name: A linear diubiquitin-based probe for efficient and selective detection of the deubiquitinating enzyme OTULIN