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Mol Cell Proteomics. 2018 Jul;17(7):1410-1425. doi: 10.1074/mcp.RA118.000676. Epub 2018 Apr 18.

A Selective Extracellular Matrix Proteomics Approach Identifies Fibronectin Proteolysis by A Disintegrin-like and Metalloprotease Domain with Thrombospondin Type 1 Motifs (ADAMTS16) and Its Impact on Spheroid Morphogenesis.

Author information

1
From the ‡Friedrich Miescher Institute for Biomedical Research, Basel, Switzerland; aptes@ccf.org rahel.schnellmann@fmi.ch.
2
§Faculty of Science, University of Basel, Basel, Switzerland.
3
¶Department of Biomedical Engineering (ND20), Cleveland Clinic Lerner Research Institute, 9500 Euclid Avenue, Cleveland, Ohio 44195.
4
From the ‡Friedrich Miescher Institute for Biomedical Research, Basel, Switzerland.
5
‖Department of Biomolecular Chemistry, University of Wisconsin, Madison, Wisconsin.
6
¶Department of Biomedical Engineering (ND20), Cleveland Clinic Lerner Research Institute, 9500 Euclid Avenue, Cleveland, Ohio 44195; aptes@ccf.org rahel.schnellmann@fmi.ch.

Abstract

Secreted and cell-surface proteases are major mediators of extracellular matrix (ECM) turnover, but their mechanisms and regulatory impact are poorly understood. We developed a mass spectrometry approach using a cell-free ECM produced in vitro to identify fibronectin (FN) as a novel substrate of the secreted metalloprotease ADAMTS16. ADAMTS16 cleaves FN between its (I)5 and (I)6 modules, releasing the N-terminal 30 kDa heparin-binding domain essential for FN self-assembly. ADAMTS16 impairs FN fibrillogenesis as well as fibrillin-1 and tenascin-C assembly, thus inhibiting formation of a mature ECM by cultured fibroblasts. Furthermore ADAMTS16 has a marked morphogenetic impact on spheroid formation by renal tubule-derived MDCKI cells. The N-terminal FN domain released by ADAMTS16 up-regulates MMP3, which cleaves the (I)5-(I)6 linker of FN similar to ADAMTS16, therefore creating a proteolytic feed-forward mechanism. Thus, FN proteolysis not only regulates FN turnover, but also FN assembly, with potential long-term consequences for ECM assembly and morphogenesis.

KEYWORDS:

ADAMTS protease; Extracellular matrix*; Fibronectin; Metalloprotease; Post-translational modifications*; Proteases*; Protein Degradation*; Proteolysis*

PMID:
29669734
PMCID:
PMC6030725
DOI:
10.1074/mcp.RA118.000676
[Indexed for MEDLINE]
Free PMC Article

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