A substrate-optimized electrophoretic mobility shift assay for ADAM12

Research output: Contribution to journalJournal articleResearchpeer-review

  • Alexander Kotzsch
  • Tine Skovgaard
  • Uwe Buus
  • Simon Andersen
  • Kanchan Devkota
  • Jens Berthelsen

ADAM12 belongs to the A disintegrin and metalloprotease (ADAM) family of secreted sheddases activating extracellular growth factors such as epidermal growth factor receptor (EGFR) ligands and tumor necrosis factor-alpha (TNF-α). ADAM proteases, most notably ADAM17 (TNF-α-converting enzyme), have long been investigated as pharmaceutical drug targets; however, due to lack of potency and in vivo side effects, none of the small-molecule inhibitors discovered so far has made it beyond clinical testing. Ongoing research on novel selective inhibitors of ADAMs requires reliable biochemical assays to validate molecular probes from large-scale screening efforts. Here we describe an electrophoretic mobility shift assay for ADAM12 based on the identification of an optimized peptide substrate that is characterized by excellent performance and reproducibility.

Original languageEnglish
JournalAnalytical Biochemistry
Pages (from-to)34-42
Number of pages9
Publication statusPublished - 1 May 2014

ID: 117612044