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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
Rahel Schnellmann, Ragna Sack, Daniel Hess, Douglas S. Annis, Deane F. Mosher, Suneel S. Apte, and Ruth Chiquet-Ehrismann.
Molecular & Cellular Proteomics, 2018, 17, 1410-1425
The authors investigated the role of the secreted protease ADAMTS16 and its effect on fibronectin (FN) and the assembly of the extra cellular matrix (ECM). Until now ADAMTS16 had been an orphan protease, without known substrates, although it has been linked to several human diseases such as cancer and hypertension.
They used a cell-free ECM produced in vitro by BALB/c fibroblasts and seeded it with HEK-EBNA cells that were transfected to produce the ADAMTS16 protease. HEK-EBNA cells stably expressing various ADAMTS16 constructs were seeded on top of the FN and the medium was analyzed by Western blotting and LC-MS/MS.
The authors confirmed a large reduction of FN fibrils in the presence of ADAMTS16, and temporal analysis of the FN assembly suggests that ADAMTS16 primarily interferes with initiation and maturation of the fibrils. Additionally the release of the 30 kDa FN fragment indicates that ADAMTS16 is also capable of degrading FN fibrils.
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