Dataset Information

The Identification of Proteolytic Substrates of Calpain-5 with N-terminomics

ABSTRACT: 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.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Daniel Young

LAB HEAD: Antoine Dufour

PROVIDER: PXD064313 | Pride | 2025-07-21

REPOSITORIES: Pride

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Dataset's files

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			Action	DRS
	S10407_27_Gabe_PreTails_1_200226.raw	Raw
	S10408_28_Gabe_PreTails_2_200226.raw	Raw
	S10409_29_Gabe_PreTails_3_200226.raw	Raw
	S10410_30_Gabe_PreTails_4_200226.raw	Raw
	S10411_31_Gabe_Tails_1_200226.raw	Raw

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The Identification of Proteolytic Substrates of Calpain-5 with N-Terminomics.

Gal Jozsef J Dufour Antoine A Young Daniel D Yang Eddy S ES Geddes James W JW

International journal of molecular sciences 20250704 13

Calpain-5/CAPN5 is a calcium-activated, non-lysosomal cysteine (thiol) protease. The substrate repertoire of CAPN5 is not known. Calpains catalyze limited proteolysis of their substrates, generating neo-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 approach called TAILS (Terminal amine isotopic labeling of substrates) to quantitatively compare the N-terminal peptides detected in ...[more]

PMID: 40650235

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