title: Development of an Assay for Measuring Proprotein-Conversion Activity on a Multiplex Magnetic Bead-Based Array Platform.
abstract: This chapter describes a protocol for measuring prolyl oligopeptidase (POP) activity using a biotinylated peptide substrate coupled to magnetic microspheres. The complex is incubated in the presence of a pituitary extract and activity can be detected by loss of the biotin label. The assay can be multiplexed for measuring multiple proprotein-cleaving enzymes simultaneously and can be used in analyses of neuropsychiatric diseases in which proteolytic cleavage of biologically active peptides is known to play a role.
authors.affiliation: Laboratory of Neuroproteomics, Institute of Biology, University of Campinas, Cidade Universitária Zeferino Vaz, 13083-862, Campinas, SP, Brazil. paulcguest@yahoo.com.
authors.forename: Paul C
authors.initials: PC
authors.lastname: Guest
authors.affiliation: Department of Chemical Engineering and Biotechnology, University of Cambridge, Cambridge, UK.
authors.forename: Divya
authors.initials: D
authors.lastname: Krishnamurthy
authors.affiliation: Department of Chemical Engineering and Biotechnology, University of Cambridge, Cambridge, UK.
authors.forename: Hassan
authors.initials: H
authors.lastname: Rahmoune
mesh_terms: D019921:Biotinylation; D006801:Humans; D018189:Immunomagnetic Separation / Q000379:methods*; D008863:Microspheres; D009419:Nerve Tissue Proteins / Q000032:analysis*; D010446:Peptide Fragments / Q000737:chemistry; D010902:Pituitary Gland / Q000201:enzymology*; D000085822:Prolyl Oligopeptidases; D012559:Schizophrenia / Q000378:metabolism; D012697:Serine Endopeptidases / Q000032:analysis*; D019809:Streptavidin
chemical_list: D009419:Nerve Tissue Proteins; D010446:Peptide Fragments; D019809:Streptavidin; D012697:Serine Endopeptidases; C000705287:PREPL protein, human; D000085822:Prolyl Oligopeptidases
keywords: Bioactive peptides; Magnetic beads; Multiplex microspheres; Prolyl oligopeptidase; Proprotein-converting enzyme; Proteomics