PXD042358-1

PXD042358 is an original dataset announced via ProteomeXchange.

Title	Identification of membrane proteins regulated by ADAM15 by SUSPECS proteomics
Description	ADAM15 is a member of the disintegrin-metalloproteinase family of sheddases, which plays a role in several biological processes including cartilage homeostasis. In contrast with well-characterized ADAMs, such as the canonical sheddases ADAM17 and ADAM10, little is known about substrates of ADAM15 or how the enzyme exerts its biological functions. Herein, we used “surface-spanning enrichment with click-sugars (SUSPECS)” proteomics to identify ADAM15 substrates and/or proteins regulated by the proteinase at the cell surface of chondrocyte-like cells. Silencing of ADAM15 by siRNAs significantly altered membrane levels of 13 proteins, all previously not known to be regulated by ADAM15. We used orthogonal techniques to validate ADAM15 effects on 3 of these proteins which have known roles in cartilage homeostasis. This confirmed that ADAM15-silencing increased cell surface levels of the programmed cell death 1 ligand 2 (PDCD1LG2) and reduced cell surface levels of vasorin and the sulfate transporter SLC26A2 through an unknown post-translational mechanism. The increase of PDCD1LG2 by ADAM15 knockdown, a single-pass type I transmembrane protein, suggested it could be a proteinase substrate. However, shed PDCD1LG2 could not be detected even by a data-independent acquisition mass spectrometry, a highly sensitive method for identification and quantification of proteins in complex protein samples, suggesting that ADAM15 regulates PDCD1LG2 membrane levels by a mechanism different from ectodomain shedding.
HostingRepository	PRIDE
AnnounceDate	2023-06-13
AnnouncementXML	Submission_2023-06-13_06:16:32.643.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	MatteoCalligaris
SpeciesList	scientific name: Homo sapiens (Human); NCBI TaxID: 9606;
ModificationList	monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Orbitrap Exploris 480

Revision	Datetime	Status	ChangeLog Entry
0	2023-05-19 05:50:49	ID requested
⏵ 1	2023-06-13 06:16:33	announced
2	2023-11-14 08:57:21	announced	2023-11-14: Updated project metadata.

submitter keyword: chondrocytes, proteomics, osteoarthritis, mass spectrometry-LC-MS/MS,Adams, SPECS, ADAM15, metalloproteases

Simone DarioScilabra
contact affiliation	Fondazione Ri.MED
contact email	sdscilabra@fondazionerimed.com
lab head
MatteoCalligaris
contact affiliation	Fondazione Ri.MED
contact email	mcalligaris@fondazionerimed.com
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD042358
     1. Label: PRIDE project
     2. Name: Identification of membrane proteins regulated by ADAM15 by SUSPECS proteomics