PXD030381-1

PXD030381 is an original dataset announced via ProteomeXchange.

Title	Mechanism-based traps enable protease and hydrolase substrate discovery
Description	we report a strategy for creating mechanism-based, light activated protease and hydrolase substrate traps in complex mixtures and live mammalian cells. The traps capture substrates of hydrolases, which normally use a serine or cysteine nucleophile. Replacing the catalytic nucleophile with genetically encoded 2,3-diaminopropionic acid permits the first step reaction to form an acyl-enzyme intermediate in which a substrate fragment is covalently linked to the enzyme through a stable amide bond; this enables stringent purification and identification of substrates. We identify new substrates for proteases, including an intramembrane mammalian rhomboid protease RHBDL4. We demonstrate that RHBDL4 can shed luminal fragments of ER-resident type I transmembrane proteins to the extracellular space, as well as catalysing non-canonical secretion of endogenous soluble ER-resident chaperones. We also discover that the putative serine hydrolase retinoblastoma binding protein 9 is an aminopeptidase – with a preference for removing aromatic amino acids – in human cells. Our results exemplify a powerful paradigm for discovering the substrates and activities of hydrolase enzymes.
HostingRepository	PRIDE
AnnounceDate	2022-02-10
AnnouncementXML	Submission_2022-02-10_09:17:01.112.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Shan Tang
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	iodoacetamide derivatized residue; deamidated residue
Instrument	Q Exactive HF; LTQ Orbitrap Velos; Orbitrap Fusion

Revision	Datetime	Status	ChangeLog Entry
0	2021-12-14 01:26:18	ID requested
⏵ 1	2022-02-10 09:17:01	announced

Dataset with its publication pending

submitter keyword: human, LC-MS/MS

Jason W. Chin
contact affiliation	Medical Research Council Laboratory of Molecular Biology
contact email	chin@mrc-lmb.cam.ac.uk
lab head
Shan Tang
contact affiliation	MRC-LMB
contact email	stang@mrc-lmb.cam.ac.uk
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD030381
     1. Label: PRIDE project
     2. Name: Mechanism-based traps enable protease and hydrolase substrate discovery