Proteolysis-induced N-terminal ectodomain shedding of the integral membrane glycoprotein CUB domain-containing protein 1 (CDCP1) is accompanied by tyrosine phosphorylation of its C-terminal domain and recruitment of Src and PKC?

He, Yaowu, Wortmann, Andreas, Burke, Leslie, Reid, Janet, Adams, Mark, Abdul-Jabbar, Ibtissam, Quigley, James, Leduc, Richard, Kirchhofer, Daniel, & Hooper, John (2010) Proteolysis-induced N-terminal ectodomain shedding of the integral membrane glycoprotein CUB domain-containing protein 1 (CDCP1) is accompanied by tyrosine phosphorylation of its C-terminal domain and recruitment of Src and PKC? Journal of Biological Chemistry, 285(34), pp. 26162-26173.

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Abstract

CUB-domain-containing protein 1 (CDCP1) is an integral membrane glycoprotein with potential as a marker and therapeutic target for a number of cancers. Here we examine mechanisms regulating cellular processing of CDCP1. By analyzing cell lines exclusively passaged non-enzymatically and through use of a panel of protease inhibitors, we demonstrate that full-length 135 kDa CDCP1 is post-translationally processed in a range of cell lines by a mechanism involving serine protease activity, generating a C-terminal 70-kDa fragment. Immunopurification and N-terminal sequencing of this cell-retained fragment and detailed mutagenesis, show that proteolytic processing of CDCP1 occurs at two sites, Arg-368 and Lys-369. We show that the serine protease matriptase is an efficient, but not essential, cellular processor of CDCP1 at Arg-368. Importantly, we also demonstrate that proteolysis induces tyrosine phosphorylation of 70-kDa CDCP1 and recruitment of Src and PKCδ to this fragment. In addition, Western blot and mass spectroscopy analyses show that an N-terminal 65-kDa CDCP1 ectodomain is shed intact from the cell surface. These data provide new insights into mechanisms regulating CDCP1 and suggest that the biological role of this protein and, potentially, its function in cancer, may be mediated by both 70-kDa cell retained and 65-kDa shed fragments, as well as the full-length 135-kDa protein.

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42 citations in Scopus
38 citations in Web of Science®
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ID Code: 42909
Item Type: Journal Article
Refereed: Yes
Additional Information: This research was originally published in the Journal of Biological Chemistry. Yaowu He, Andreas Wortmann, Les J. Burke, Janet C. Reid, Mark N. Adams, Ibtissam Abdul-Jabbar, James P. Quigley, Richard Leduc, Daniel Kirchhofer and John D. Hooper. Proteolysis-induced N-terminal Ectodomain Shedding of the Integral Membrane Glycoprotein CUB Domain-containing Protein 1 (CDCP1) Is Accompanied by Tyrosine Phosphorylation of Its C-terminal Domain and Recruitment of Src and PKCδ. Journal of Biological Chemistry. 2010; 285:26162-26173. © the American Society for Biochemistry and Molecular Biology.
Keywords: Membrane Proteins, Phosphotyrosine, Protease, Receptors, Shedding, CDCP1, PKC(delta), Src, Serine Protease, Tyrosine Phosphorylation
DOI: 10.1074/jbc.M109.096453
ISSN: 0021-9258
Subjects: Australian and New Zealand Standard Research Classification > BIOLOGICAL SCIENCES (060000) > BIOCHEMISTRY AND CELL BIOLOGY (060100)
Australian and New Zealand Standard Research Classification > ENGINEERING (090000) > CHEMICAL ENGINEERING (090400)
Divisions: Current > QUT Faculties and Divisions > Faculty of Health
Past > QUT Faculties & Divisions > Faculty of Science and Technology
Current > Institutes > Institute of Health and Biomedical Innovation
Deposited On: 13 Jul 2011 13:08
Last Modified: 14 May 2017 23:00

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