title: Functional proteomics of kallikrein-related peptidases in ovarian cancer ascites fluid.
abstract: Kallikrein-related peptidases (KLKs) are secreted serine proteinases with trypsin or chymotrypsin-like activity. Several family members, such as KLKs 6 and 10, are potential ovarian cancer biomarkers. Recently, using a newly developed assay for active KLK6, we found that only a very small proportion of immunoreactive KLK6 in tumor-derived clinical samples (malignant ascites fluid), in cerebrospinal fluid, and in cancer cell line supernatants is enzymatically active. We therefore hypothesized that a proportion of other immunoreactive KLKs in such samples could be present, but might be partly complexed to endogenous serine proteinase inhibitors. Using a combination of immunological isolation of the enzymes, activity-based probe analysis and proteomics, we identified active KLK10 in ovarian cancer ascites and we provide preliminary data that the activity of other KLKs present in these samples can be decreased by known proteinase inhibitors (e.g., alpha2-macroglobulin, alpha1-antitrypsin). Our data suggest that the enzymatic activity of ovarian cancer-released KLKs that are detected by regular immunoassays is low in vivo and very likely regulated by proteinase inhibitors.
authors.affiliation: Department of Physiology and Pharmacology, University of Calgary, Alberta, Canada.
authors.forename: Katerina
authors.initials: K
authors.lastname: Oikonomopoulou
authors.forename: Ihor
authors.initials: I
authors.lastname: Batruch
authors.forename: Chris R
authors.initials: CR
authors.lastname: Smith
authors.forename: Antoninus
authors.initials: A
authors.lastname: Soosaipillai
authors.forename: Eleftherios P
authors.initials: EP
authors.lastname: Diamandis
authors.forename: Morley D
authors.initials: MD
authors.lastname: Hollenberg
mesh_terms: D001201:Ascites / Q000201:enzymology*; D002850:Chromatography, Gel; D005260:Female; D006801:Humans; D007610:Kallikreins / Q000737:chemistry / Q000378:metabolism*; D015335:Molecular Probes / Q000378:metabolism; D008970:Molecular Weight; D010051:Ovarian Neoplasms / Q000201:enzymology*; D011480:Protease Inhibitors / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D014357:Trypsin / Q000378:metabolism
chemical_list: D015335:Molecular Probes; D011480:Protease Inhibitors; D007610:Kallikreins; D014357:Trypsin