Substrate-biased activity-based probes identify proteases that cleave receptor CDCP1

Thomas Kryza, Tashbib Khan, Scott Lovell, Brittney S Harrington, Julia Yin, Sean Porazinski, Marina Pajic, Hannu Koistinen, Juha K Rantala, Tobias Dreyer, Viktor Magdolen, Ute Reuning, Yaowu He, Edward W Tate, John D Hooper

Research output: Contribution to journalArticlepeer-review

16 Citations (SciVal)

Abstract

CUB domain-containing protein 1 (CDCP1) is an oncogenic orphan transmembrane receptor and a promising target for the detection and treatment of cancer. Extracellular proteolysis of CDCP1 by poorly defined mechanisms induces pro-metastatic signaling. We describe a new approach for the rapid identification of proteases responsible for key proteolytic events using a substrate-biased activity-based probe (sbABP) that incorporates a substrate cleavage motif grafted onto a peptidyl diphenyl phosphonate warhead for specific target protease capture, isolation and identification. Using a CDCP1-biased probe, we identify urokinase (uPA) as the master regulator of CDCP1 proteolysis, which acts both by directly cleaving CDCP1 and by activating CDCP1-cleaving plasmin. We show that coexpression of uPA and CDCP1 is strongly predictive of poor disease outcome across multiple cancers and demonstrate that uPA-mediated CDCP1 proteolysis promotes metastasis in disease-relevant preclinical in vivo models. These results highlight CDCP1 cleavage as a potential target to disrupt cancer and establish sbABP technology as a new approach to identify disease-relevant proteases.

Original languageEnglish
Pages (from-to)776-783
Number of pages8
JournalNature Chemical Biology
Volume17
Issue number7
Early online date15 Apr 2021
DOIs
Publication statusPublished - 31 Jul 2021

Keywords

  • Animals
  • Antigens, Neoplasm/chemistry
  • Cell Adhesion Molecules/chemistry
  • Humans
  • Mice
  • Mice, Inbred NOD
  • Mice, SCID
  • Molecular Structure
  • Peptide Hydrolases/analysis
  • Substrate Specificity

Fingerprint

Dive into the research topics of 'Substrate-biased activity-based probes identify proteases that cleave receptor CDCP1'. Together they form a unique fingerprint.

Cite this