Development of a novel assay for proprotein converting enzyme activity on a multiplex bead‐based array system. Issue 20 (8th October 2013)

• Record Type: Journal Article
• Authors: Albrecht, Andreas; Rahmoune, Hassan; Leedjärv, Kristi; Knorpp, Thomas; Joos, Thomas; Stocki, Pawel; Guest, Paul C.; Bahn, Sabine
• Is Part Of: Proteomics. Volume 13:Issue 20(2013:Oct.)
• Page Start: 2976
• Page End: 2979
• Publication Date: 2013-10-08
• Subjects: Proteins -- Separation -- Periodicals; Bioinformatics -- Periodicals; Proteomics -- Periodicals; Genomes -- Periodicals; Molecular genetics -- Periodicals; 572.605
• DOI: 10.1002/pmic.201300060
• Languages: English
• ISSNs: 1615-9853

Abstract

<abstract abstract-type="main"> <title> <x xml:space="preserve">Abstract</x> </title> <p>We describe the development of a novel, robust assay system for determining the changes in activity of proprotein converting enzymes. An assay for prolyl oligopeptidase (POP) activity was constructed using a peptide‐streptavidin substrate coupled to magnetic microspheres and cleavage was detected by loss of streptavidin on the MAGPIX reader. Test analysis of postmortem pituitary extracts from schizophrenia patients showed an increase in POP activity compared to controls. The results were validated using both fluorometric and Western blot analyses for POP activity and immunoreactivity, respectively. The assays can be multiplexed for measuring the activity of multiple proprotein cleaving enzymes simultaneously in laboratory and clinical settings and should add valuable new information for conditions such as neuropsychiatric diseases, diabetes, endocrine dysfunction, and cancer, where effects on proteolysis of biologically active peptides play a key role.</p> </abstract>