The substrate degradome of meprin metalloproteases reveals an unexpected proteolytic link between meprin β and ADAM10

Tamara Jefferson, Ulrich Auf Dem Keller, Caroline Bellac, Verena V. Metz, Claudia Broder, Jana Hedrich, Anke Ohler, Wladislaw Maier, Viktor Magdolen, Erwin Sterchi, Judith S. Bond, Arumugam Jayakumar, Heiko Traupe, Athena Chalaris, Stefan Rose-John, Claus U. Pietrzik, Rolf Postina, Christopher M. Overall, Christoph Becker-Pauly*

*Corresponding author for this work

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

The in vivo roles of meprin metalloproteases in pathophysiological conditions remain elusive. Substrates define protease roles. Therefore, to identify natural substrates for human meprin α and β we employed TAILS (terminal amine isotopic labeling of substrates), a proteomics approach that enriches for N-terminal peptides of proteins and cleavage fragments. Of the 151 new extracellular substrates we identified, it was notable that ADAM10 (a disintegrin and metalloprotease domain-containing protein 10)-the constitutive α-secretase-is activated by meprin β through cleavage of the propeptide. To validate this cleavage event, we expressed recombinant proADAM10 and after preincubation with meprin β, this resulted in significantly elevated ADAM10 activity. Cellular expression in murine primary fibroblasts confirmed activation. Other novel substrates including extracellular matrix proteins, growth factors and inhibitors were validated by western analyses and enzyme activity assays with Edman sequencing confirming the exact cleavage sites identified by TAILS. Cleavages in vivo were confirmed by comparing wild-type and meprin-/- mice. Our finding of cystatin C, elafin and fetuin-A as substrates and natural inhibitors for meprins reveal new mechanisms in the regulation of protease activity important for understanding pathophysiological processes.

Original languageEnglish
JournalCellular and Molecular Life Sciences
Volume70
Issue number2
Pages (from-to)309-333
ISSN1420-682X
DOIs
Publication statusPublished - 2013
Externally publishedYes

Keywords

  • ADAM10
  • Degradome
  • Meprin
  • Metalloproteases
  • Proteomics
  • TAILS

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