PXD001643

PXD001643 is an original dataset announced via ProteomeXchange.

Title	SILAC-iTRAQ-TAILS - Monitoring matrix metalloproteinase activity at the epidermal-dermal interface by SILAC-iTRAQ-TAILS
Description	Secreted proteases act on interstitial tissue secretomes released from multiple cell types. Thus, substrate proteins might be part of higher molecular complexes constituted by many proteins with diverse and potentially unknown cellular origin. In cell culture these might be reconstituted by mixing native secretomes from different cell types prior to incubation with a test protease. Although current degradomics techniques could identify novel substrate proteins in these complexes, all information on the cellular origin would be lost. To address this limitation we combined iTRAQ-based Terminal Amine Isotopic Labeling of Substrates (iTRAQ-TAILS) with stable isotope labeling by amino acids in cell culture (SILAC) to assign proteins to a specific cell type by MS1- and their cleavage by MS2-based quantification in the same experiment. We demonstrate the power of our newly established workflow by monitoring matrix metalloproteinase (MMP) 10-dependent cleavages in mixtures from heavy labeled fibroblast and light labeled keratinocyte sectretomes. This analysis correctly assigned extracellular matrix components, such as laminins and collagens, to their respective cellular origins and revealed their processing in an MMP10-dependent manner. Hence, our newly devised degradomics workflow facilitates deeper insights into protease activity in complex intercellular compartments like the epidermal-dermal interface by integrating multiple modes of quantification with positional proteomics.
HostingRepository	PRIDE
AnnounceDate	2024-10-22
AnnouncementXML	Submission_2024-10-22_04:22:16.503.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Ulrich auf dem Keller
SpeciesList	scientific name: Mus musculus (Mouse); NCBI TaxID: 10090;
ModificationList	6x(13)C; iTRAQ4plex-116 reporter+balance reagent acylated residue; acetylated residue; monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2015-01-02 03:11:45	ID requested
1	2015-05-11 08:01:59	announced
⏵ 2	2024-10-22 04:22:18	announced	2024-10-22: Updated project metadata.

10.1002/pmic.201400627;

Schlage P, Kockmann T, Kizhakkedathu JN, auf dem Keller U, Monitoring matrix metalloproteinase activity at the epidermal-dermal interface by SILAC-iTRAQ-TAILS. Proteomics, 15(14):2491-502(2015) [pubmed]

curator keyword: Technical, Biological

submitter keyword: skin, basement membrane, TAILS,SILAC, iTRAQ, MMP

Ulrich auf dem Keller
contact affiliation	ETH Zurich, Institute of Molecular Health Sciences, Zurich, Switzerland
contact email	ulrich.aufdemkeller@biol.ethz.ch
lab head
Ulrich auf dem Keller
contact affiliation	Technical University of Denmark
contact email	ulrich.aufdemkeller@biol.ethz.ch
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/2015/05/PXD001643

PRIDE project URI

• PRIDE
  1. PXD001643
     1. Label: PRIDE project
     2. Name: SILAC-iTRAQ-TAILS - Monitoring matrix metalloproteinase activity at the epidermal-dermal interface by SILAC-iTRAQ-TAILS