Biochemical and functional characterization of the human tissue kallikrein 9
Autor: | Davor Brinc, Yijing Yu, Eleftherios P. Diamandis, Ioannis Prassas, Panagiota Filippou, Sofia Farkona |
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Rok vydání: | 2017 |
Předmět: |
0301 basic medicine
Proteases Glycosylation Serine Proteinase Inhibitors medicine.medical_treatment Tissue kallikrein Biology Biochemistry Cell Line Substrate Specificity 03 medical and health sciences Catalytic Domain Cell Line Tumor Enzyme Stability medicine Humans Magnesium Nerve Growth Factors Tyrosine Molecular Biology chemistry.chemical_classification Protease Midkine Cell Biology Kallikrein Enzyme assay Peptide Fragments Recombinant Proteins Cell biology HaCaT Kinetics Zinc 030104 developmental biology Enzyme HEK293 Cells chemistry Proteolysis biology.protein Calcium Kallikreins Protein Processing Post-Translational |
Zdroj: | The Biochemical journal. 474(14) |
ISSN: | 1470-8728 |
Popis: | Human tissue kallikrein 9 (KLK9) is a member of the kallikrein-related family of proteases. Despite its known expression profile, much less is known about the functional roles of this protease and its implications in normal physiology and disease. We present here the first data on the biochemical characterization of KLK9, investigate parameters that affect its enzymatic activity (such as inhibitors) and provide preliminary insights into its putative substrates. We show that mature KLK9 is a glycosylated chymotrypsin-like enzyme with strong preference for tyrosine over phenylalanine at the P1 cleavage position. The enzyme activity is enhanced by Mg2+ and Ca2+, but is reversibly attenuated by Zn2+. KLK9 is inhibited in vitro by many naturally occurring or synthetic protease inhibitors. Using a combination of degradomic and substrate specificity assays, we identified candidate KLK9 substrates in two different epithelial cell lines [the non-tumorigenic human keratinocyte cells (HaCaT) and the tumorigenic tongue squamous carcinoma cells (SCC9)]. Two potential KLK9 substrates [KLK10 and midkine (MDK)] were subjected to further validation. Taken together, our data delineate some functional and biochemical properties of KLK9 for future elucidation of the role of this enzyme in health and disease. |
Databáze: | OpenAIRE |
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