Journal article
The substrate degradome of meprin metalloproteases reveals an unexpected proteolytic link between meprin β and ADAM10
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.
Language: | English |
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Publisher: | SP Birkhäuser Verlag Basel |
Year: | 2013 |
Pages: | 309-33 |
ISSN: | 14209071 and 1420682x |
Types: | Journal article |
DOI: | 10.1007/s00018-012-1106-2 |
ADAM Proteins ADAM10 ADAM10 Protein ADAM10 protein, human Amino Acid Sequence Amyloid Precursor Protein Secretases Animals Biochemistry, general Biomedical and Life Sciences Biomedicine general Caco-2 Cells Cell Biology Cell Line Cystatin C Cytokines Degradome Elafin Extracellular Matrix Proteins HEK293 Cells Humans Life Sciences Life Sciences, general Membrane Proteins Meprin Metalloendopeptidases Metalloproteases Mice Mice, Knockout Proteomics TAILS alpha-2-HS-Glycoprotein meprin A