PXD064313

PXD064313 is an original dataset announced via ProteomeXchange.

Title	The Identification of Proteolytic Substrates of Calpain-5 with N-terminomics
Description	Calpain-5/CAPN5 is a calcium-activated, non-lysosomal cysteine (thiol) protease. The substrate spectrum repertoire of CAPN5 is not known. Calpains catalyze limited proteolysis of their substrates, generating novel neo protein N-termini that correspond to internal residues of their nascent substrate proteins. To identify such neo-N-termini generated by CAPN5, we employed an N-terminomics approached called the TAILS (Terminal amine isotopic labeling of substrates) proteomics method to quantitatively compare the N-terminal peptides detected in parental and CAPN5-deficient SH-SY5Y neuroblastoma cells. Thirty neo-N-termini corresponding to 29 protein groups and 24 unique proteins were found detected to be depleted in the CAPN5-/- cells. A subset of the identified putative substrates was further studied with CAPN5 co-immunoprecipitation, in vitro calcium-induced CAPN5 proteolysis assay, and comparing their cellular fragmentation patterns in parental and CAPN5-deficient SH-SY5Y cells. Here, wWe provide evidence for CAPN5-mediated proteolysis of the synaptic proteins DLGAP4, IQSEC1 and MPDZ, the neurodegeneration-related EWS, hnRNPU, TFG and UGP2, the DNA replication regulator MCM3, and the neuronal differentiation regulator LMTK1. Our data provide newThe relevance of the newly identified CAPN5 substrates tofor Neovascular neovascular inflammatory vitreoretinopathy (NIV), a progressive eye disease caused by pathogenic mutations in CAPN5, is discussed.
HostingRepository	PRIDE
AnnounceDate	2025-07-21
AnnouncementXML	Submission_2025-07-20_16:09:06.864.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Daniel Young
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	acetylated residue; iodoacetamide derivatized residue
Instrument	Orbitrap Fusion Lumos

Revision	Datetime	Status	ChangeLog Entry
0	2025-05-26 15:34:21	ID requested
⏵ 1	2025-07-20 16:09:07	announced

10.3390/ijms26136459;

Gal J, Dufour A, Young D, Yang ES, Geddes JW, The Identification of Proteolytic Substrates of Calpain-5 with N-Terminomics. Int J Mol Sci, 26(13):(2025) [pubmed]

submitter keyword: CAPN5, protease, disease, TAILS, N-terminomics,Calpain-5

Antoine Dufour
contact affiliation	Department of Physiology and Pharmacology, Cumming School of Medicine, University of Calgary, Alberta T2N 1N4 Canada Snyder Insitute for Chronic Diseases, Cumming School of Medicine, University of Calgary, Alberta T2N 1N4 Canada
contact email	Antoine.dufour@ucalgary.ca
lab head
Daniel Young
contact affiliation	University of Calgary
contact email	Daniel.young1@ucalgary.ca
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2025/07/PXD064313

PRIDE project URI

• PRIDE
  1. PXD064313
     1. Label: PRIDE project
     2. Name: The Identification of Proteolytic Substrates of Calpain-5 with N-terminomics