title: Determination of the substrate repertoire of ADAMTS2, 3, and 14 significantly broadens their functions and identifies extracellular matrix organization and TGF-β signaling as primary targets.
abstract: A disintegrin and metalloproteinase with thrombospondin type I motif (ADAMTS)2, 3, and 14 are collectively named procollagen N-proteinases (pNPs) because of their specific ability to cleave the aminopropeptide of fibrillar procollagens. Several reports also indicate that they could be involved in other biological processes, such as blood coagulation, development, and male fertility, but the potential substrates associated with these activities remain unknown. Using the recently described N-terminal amine isotopic labeling of substrate approach, we analyzed the secretomes of human fibroblasts and identified 8, 17, and 22 candidate substrates for ADAMTS2, 3, and 14, respectively. Among these newly identified substrates, many are components of the extracellular matrix and/or proteins related to cell signaling such as latent TGF-β binding protein 1, TGF-β RIII, and dickkopf-related protein 3. Candidate substrates for the 3 ADAMTS have been biochemically validated in different contexts, and the implication of ADAMTS2 in the control of TGF-β activity has been further demonstrated in human fibroblasts. Finally, the cleavage site specificity was assessed showing a clear and unique preference for nonpolar or slightly hydrophobic amino acids. This work shows that the activities of the pNPs extend far beyond the classically reported processing of the aminopropeptide of fibrillar collagens and that they should now be considered as multilevel regulators of matrix deposition and remodeling.-Bekhouche, M., Leduc, C., Dupont, L., Janssen, L., Delolme, F., Vadon-Le Goff, S., Smargiasso, N., Baiwir, D., Mazzucchelli, G., Zanella-Cleon, I., Dubail, J., De Pauw, E., Nusgens, B., Hulmes, D. J. S., Moali, C., Colige, A. Determination of the substrate repertoire of ADAMTS2, 3, and 14 significantly broadens their functions and identifies extracellular matrix organization and TGF-β signaling as primary targets.
authors.affiliation: Laboratory of Connective Tissues Biology, University of Liège, Liège, Belgium; mouradbekhouche2@gmail.com.
authors.forename: Mourad
authors.initials: M
authors.lastname: Bekhouche
authors.affiliation: Laboratory of Connective Tissues Biology, University of Liège, Liège, Belgium;
authors.forename: Cedric
authors.initials: C
authors.lastname: Leduc
authors.affiliation: Laboratory of Connective Tissues Biology, University of Liège, Liège, Belgium;
authors.forename: Laura
authors.initials: L
authors.lastname: Dupont
authors.affiliation: Laboratory of Connective Tissues Biology, University of Liège, Liège, Belgium;
authors.forename: Lauriane
authors.initials: L
authors.lastname: Janssen
authors.affiliation: Tissue Biology and Therapeutic Engineering, Centre National de la Recherche Scientifique/University of Lyon Unité Mixte de Recherche 5305, Lyon, France; and Protein Science Facility, Institute for the Biology and Chemistry of Proteins, Unité Mixte de Service 3444, Lyon, France.
authors.forename: Frederic
authors.initials: F
authors.lastname: Delolme
authors.affiliation: Tissue Biology and Therapeutic Engineering, Centre National de la Recherche Scientifique/University of Lyon Unité Mixte de Recherche 5305, Lyon, France; and.
authors.forename: Sandrine
authors.initials: S
authors.lastname: Vadon-Le Goff
authors.affiliation: Mass Spectrometry Laboratory, GIGA Proteomics, University of Liège, Liège, Belgium;
authors.forename: Nicolas
authors.initials: N
authors.lastname: Smargiasso
authors.affiliation: GIGA Proteomic Facility, GIGA-Interdisciplinary Cluster for Applied Genoproteomics, University of Liège, Liège, Belgium;
authors.forename: Dominique
authors.initials: D
authors.lastname: Baiwir
authors.affiliation: Mass Spectrometry Laboratory, GIGA Proteomics, University of Liège, Liège, Belgium;
authors.forename: Gabriel
authors.initials: G
authors.lastname: Mazzucchelli
authors.affiliation: Protein Science Facility, Institute for the Biology and Chemistry of Proteins, Unité Mixte de Service 3444, Lyon, France.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Zanella-Cleon
authors.affiliation: Laboratory of Connective Tissues Biology, University of Liège, Liège, Belgium;
authors.forename: Johanne
authors.initials: J
authors.lastname: Dubail
authors.affiliation: Mass Spectrometry Laboratory, GIGA Proteomics, University of Liège, Liège, Belgium;
authors.forename: Edwin
authors.initials: E
authors.lastname: De Pauw
authors.affiliation: Laboratory of Connective Tissues Biology, University of Liège, Liège, Belgium;
authors.forename: Betty
authors.initials: B
authors.lastname: Nusgens
authors.affiliation: Tissue Biology and Therapeutic Engineering, Centre National de la Recherche Scientifique/University of Lyon Unité Mixte de Recherche 5305, Lyon, France; and.
authors.forename: David J S
authors.initials: DJ
authors.lastname: Hulmes
authors.affiliation: Tissue Biology and Therapeutic Engineering, Centre National de la Recherche Scientifique/University of Lyon Unité Mixte de Recherche 5305, Lyon, France; and.
authors.forename: Catherine
authors.initials: C
authors.lastname: Moali
authors.affiliation: Laboratory of Connective Tissues Biology, University of Liège, Liège, Belgium; acolige@ulg.ac.be.
authors.forename: Alain
authors.initials: A
authors.lastname: Colige
mesh_terms: D000071096:ADAMTS Proteins / Q000235:genetics / Q000378:metabolism*; D048868:Adaptor Proteins, Signal Transducing; D018925:Chemokines; D005109:Extracellular Matrix / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000502:physiology; D057809:HEK293 Cells; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism; D053779:Latent TGF-beta Binding Proteins / Q000235:genetics / Q000378:metabolism; D011348:Procollagen N-Endopeptidase / Q000235:genetics / Q000378:metabolism*; D011509:Proteoglycans / Q000235:genetics / Q000378:metabolism; D018125:Receptors, Transforming Growth Factor beta / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000502:physiology; D016212:Transforming Growth Factor beta / Q000235:genetics / Q000378:metabolism*
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D018925:Chemokines; C498624:DKK3 protein, human; D036341:Intercellular Signaling Peptides and Proteins; C509032:LTBP1 protein, human; D053779:Latent TGF-beta Binding Proteins; D011509:Proteoglycans; D018125:Receptors, Transforming Growth Factor beta; D016212:Transforming Growth Factor beta; C071736:betaglycan; D000071096:ADAMTS Proteins; C447060:ADAMTS14 protein, human; C434951:ADAMTS3 protein, human; C000609307:ADAMTS2 protein, human; D011348:Procollagen N-Endopeptidase
keywords: DKK3; LTBP1; N-TAILS; betaglycan