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

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


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*.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3T3 Cells
  • ADAMTS Proteins / chemistry
  • ADAMTS Proteins / metabolism*
  • Amino Acid Sequence
  • Animals
  • Collagen / metabolism
  • Dogs
  • Extracellular Matrix / metabolism*
  • Fibroblasts / metabolism
  • Fibronectins / metabolism*
  • HEK293 Cells
  • Humans
  • Madin Darby Canine Kidney Cells
  • Matrix Metalloproteinase 3 / metabolism
  • Mice
  • Morphogenesis*
  • Peptides / chemistry
  • Peptides / metabolism
  • Protein Binding
  • Protein Domains
  • Proteolysis*
  • Proteomics / methods*
  • Spheroids, Cellular / metabolism*
  • Substrate Specificity
  • Up-Regulation


  • Fibronectins
  • Peptides
  • Collagen
  • ADAMTS Proteins
  • ADAMTS16 protein, mouse
  • Matrix Metalloproteinase 3