Biochemical characterization of human tissue kallikrein 15 and examination of its potential role in cancer

Panagiota S. Filippou, Annie H. Ren, Sudarshan Bala, Michail Dimitrios Papaioannou, Davor Brinc, Ioannis Prassas, Theano Karakosta, Eleftherios P. Diamandis

    Research output: Contribution to journalArticle

    Abstract

    Objective: Human tissue kallikrein 15 (KLK15) is the last cloned member of the KLK-related gene family. Despite being implicated in multiple cancers, its pathophysiological role remains unknown. We aimed to biochemically characterize KLK15 and preliminarily study its role in cancer. Design & methods: Recombinant KLK15 protein was produced, purified to homogeneity and quantified by mass spectrometry (parallel reaction monitoring analysis). We profiled the enzymatic activity of KLK15 using fluorogenic peptide substrates, and performed kinetic analysis to discover the cleavage sites. As KLK15 has mainly been associated with prostate cancer, we used a degradomic approach and subsequent KEGG pathway analysis to identify a number of putative protein substrates in the KLK15-treated prostate cancer cell line PC3. Results: We discovered trypsin-like activity in KLK15, finding that it cleaves preferentially after arginine (R). The enzymatic activity of KLK15 was regulated by different factors such as pH, cations and serine protease inhibitors. Notably, we revealed that KLK15 most likely interacts with the extracellular matrix (ECM) receptor group. Conclusion: To our knowledge, this is the first study that experimentally verifies the trypsin-like activity of KLK15. We show here for the first time that KLK15 may be able to cleave many ECM components, similar to several members of the KLK family. Thus the protease could potentially be linked to tumorigenesis by promoting metastasis via this mechanism.

    Original languageEnglish
    Pages (from-to)108-115
    Number of pages8
    JournalClinical Biochemistry
    Volume58
    Early online date18 Jun 2018
    DOIs
    Publication statusE-pub ahead of print - 18 Jun 2018

    Bibliographical note

    Copyright © 2018 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

    Fingerprint Dive into the research topics of 'Biochemical characterization of human tissue kallikrein 15 and examination of its potential role in cancer'. Together they form a unique fingerprint.

  • Cite this