title: Matrix Metalloproteinase 10 Degradomics in Keratinocytes and Epidermal Tissue Identifies Bioactive Substrates With Pleiotropic Functions.
abstract: Matrix metalloproteinases (MMPs) are important players in skin homeostasis, wound repair, and in the pathogenesis of skin cancer. It is now well established that most of their functions are related to processing of bioactive proteins rather than components of the extracellular matrix (ECM). MMP10 is highly expressed in keratinocytes at the wound edge and at the invasive front of tumors, but hardly any non-ECM substrates have been identified and its function in tissue repair and carcinogenesis is unclear. To better understand the role of MMP10 in the epidermis, we employed multiplexed iTRAQ-based Terminal Amine Isotopic Labeling of Substrates (TAILS) and monitored MMP10-dependent proteolysis over time in secretomes from keratinocytes. Time-resolved abundance clustering of neo-N termini classified MMP10-dependent cleavage events by efficiency and refined the MMP10 cleavage site specificity by revealing a so far unknown preference for glutamate in the P1 position. Moreover, we identified and validated the integrin alpha 6 subunit, cysteine-rich angiogenic inducer 61 and dermokine as novel direct MMP10 substrates and provide evidence for MMP10-dependent but indirect processing of phosphatidylethanolamine-binding protein 1. Finally, we sampled the epidermal proteome and degradome in unprecedented depth and confirmed MMP10-dependent processing of dermokine in vivo by TAILS analysis of epidermis from transgenic mice that overexpress a constitutively active mutant of MMP10 in basal keratinocytes. The newly identified substrates are involved in cell adhesion, migration, proliferation, and/or differentiation, indicating a contribution of MMP10 to local modulation of these processes during wound healing and cancer development. Data are available via ProteomeXchange with identifier PXD002474.
authors.affiliation: From the ‡ETH Zurich, Department of Biology,Institute of Molecular Health Sciences, Otto-Stern-Weg 7, 8093 Zurich, Switzerland;
authors.forename: Pascal
authors.initials: P
authors.lastname: Schlage
authors.affiliation: From the ‡ETH Zurich, Department of Biology,Institute of Molecular Health Sciences, Otto-Stern-Weg 7, 8093 Zurich, Switzerland;
authors.forename: Tobias
authors.initials: T
authors.lastname: Kockmann
authors.affiliation: From the ‡ETH Zurich, Department of Biology,Institute of Molecular Health Sciences, Otto-Stern-Weg 7, 8093 Zurich, Switzerland;
authors.forename: Fabio
authors.initials: F
authors.lastname: Sabino
authors.affiliation: §University of British Columbia, Department of Pathology and Laboratory Medicine and Department of Chemistry, Centre for Blood Research, 4.401Life Sciences Institute, 2350 Health Sciences Mall, Vancouver, British Columbia, Canada V6T 1Z3.
authors.forename: Jayachandran N
authors.initials: JN
authors.lastname: Kizhakkedathu
authors.affiliation: From the ‡ETH Zurich, Department of Biology,Institute of Molecular Health Sciences, Otto-Stern-Weg 7, 8093 Zurich, Switzerland; ulrich.aufdemkeller@biol.ethz.ch.
authors.forename: Ulrich
authors.initials: U
authors.lastname: Auf dem Keller
mesh_terms: D000818:Animals; D002448:Cell Adhesion; D002465:Cell Movement; D049109:Cell Proliferation; D055515:Cysteine-Rich Protein 61 / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D004817:Epidermis / Q000378:metabolism*; D005260:Female; D006801:Humans; D039503:Integrin alpha6 / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D036341:Intercellular Signaling Peptides and Proteins; D007553:Isotope Labeling; D015603:Keratinocytes / Q000378:metabolism*; D053506:Matrix Metalloproteinase 10 / Q000378:metabolism*; D051379:Mice; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D059748:Proteolysis; D020543:Proteome / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D040901:Proteomics / Q000379:methods
chemical_list: C501848:CCN1 protein, human; D055515:Cysteine-Rich Protein 61; C495568:DMKN protein, human; C588002:ITGA6 protein, human; D039503:Integrin alpha6; D036341:Intercellular Signaling Peptides and Proteins; D011506:Proteins; D020543:Proteome; C511770:MMP10 protein, human; D053506:Matrix Metalloproteinase 10