Substrate-biased activity-based probes identify proteases that cleave receptor CDCP1
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.
|ISBN||1552-4469 (Electronic) 1552-4450 (Linking)|
|Authors||Kryza, T.; Khan, T.; Lovell, S.; Harrington, B. S.; Yin, J.; Porazinski, S.; Pajic, M.; Koistinen, H.; Rantala, J. K.; Dreyer, T.; Magdolen, V.; Reuning, U.; He, Y.; Tate, E. W.; Hooper, J. D.|
|Responsible Garvan Author|
|Publisher Name||Nature Chemical Biology|
|URL link to publisher's version||https://www.ncbi.nlm.nih.gov/pubmed/33859413|