Laminin-332 is a substrate for hepsin, a protease associated with prostate cancer progression

Manish Tripathi, Srinivas Nandana, Hironobu Yamashita, Rajkumar Ganesan, Daniel Kirchhofer, Vito Quaranta

Research output: Contribution to journalArticle

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Abstract

Hepsin, a cell surface protease, is widely reported to be over-expressed in more than 90% of human prostate tumors. Hepsin expression correlates with tumor progression, making it a significant marker and target for prostate cancer. Recently, it was reported that in a prostate cancer mouse model, hepsin up-regulation in tumor tissue promotes progression and metastasis. The underlying mechanisms, however, remain largely uncharacterized. Hepsin transgenic mice displayed reduced laminin-332 (Ln-332) expression in prostate tumors. This is an intriguing cue, since proteolytic processing of extracellular matrix macromolecules, such as Ln-332, is believed to be involved in cancer progression, and Ln-332 expression is lost during human prostate cancer progression. In this study, we provide the first direct evidence that hepsin cleaves Ln-332. Cleavage is specific, since it is both inhibited in a dose-dependent manner by a hepsin inhibitor (Kunitz domain-1) and does not occur when catalytically inactive hepsin is used. By Western blotting and mass spectrometry, we determined that hepsin cleaves the β3 chain of Ln-332. N-terminal sequencing identified the cleavage site at β3 Arg245, in a sequence context (SQLR245 ↓ LQGSCFC) conserved among species and in remarkable agreement with reported consensus target sequences for hepsin activity. In vitro cell migration assays showed that hepsin-cleaved Ln-332 enhanced motility of DU145 prostate cancer cells, which was inhibited by Kunitz domain-1. Further, hepsin-overexpressing LNCaP prostate cancer cells also exhibited increased migration on Ln-332. Direct cleavage of Ln-332 may be one mechanism by which hepsin promotes prostate tumor progression and metastasis, possibly by up-regulating prostate cancer cell motility.

Original languageEnglish (US)
Pages (from-to)30576-30584
Number of pages9
JournalJournal of Biological Chemistry
Volume283
Issue number45
DOIs
StatePublished - Nov 7 2008
Externally publishedYes

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Prostatic Neoplasms
Peptide Hydrolases
Substrates
Tumors
Prostate
Neoplasms
Cells
hepsin
kalinin
Cell Migration Assays
Neoplasm Metastasis
Consensus Sequence
Macromolecules
Transgenic Mice
Cell Movement
Cues
Extracellular Matrix
Mass spectrometry
Assays
Mass Spectrometry

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Laminin-332 is a substrate for hepsin, a protease associated with prostate cancer progression. / Tripathi, Manish; Nandana, Srinivas; Yamashita, Hironobu; Ganesan, Rajkumar; Kirchhofer, Daniel; Quaranta, Vito.

In: Journal of Biological Chemistry, Vol. 283, No. 45, 07.11.2008, p. 30576-30584.

Research output: Contribution to journalArticle

Tripathi, Manish ; Nandana, Srinivas ; Yamashita, Hironobu ; Ganesan, Rajkumar ; Kirchhofer, Daniel ; Quaranta, Vito. / Laminin-332 is a substrate for hepsin, a protease associated with prostate cancer progression. In: Journal of Biological Chemistry. 2008 ; Vol. 283, No. 45. pp. 30576-30584.
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