title: Fluorescent-conjugated polymer superquenching facilitates highly sensitive detection of proteases.
abstract: Sensor formats have been developed for detecting the activity of proteolytic enzymes based on fluorescent conjugated polymer superquenching. These sensors employ a reactive peptide sequence within a tether linking a quencher to a biotin. The peptide binds to sensors containing colocated biotin-binding protein and fluorescent polymer by means of biotin-biotin binding protein interactions, resulting in a strong quenching of polymer fluorescence. Enzyme-mediated cleavage of the peptide results in a reversal of the fluorescence quenching. These assays for protease activity are simple, sensitive, fast, and have the specificity required for screening chemical libraries for novel protease inhibitors in a high-throughput screening assay environment. These assays have been demonstrated for enterokinase, caspase-3/7, and beta-secretase.
authors.affiliation: QTL Biosystems, 2778 Agua Fria Street, Santa Fe, NM 87507, USA.
authors.forename: Sriram
authors.initials: S
authors.lastname: Kumaraswamy
authors.forename: Troy
authors.initials: T
authors.lastname: Bergstedt
authors.forename: Xiaobo
authors.initials: X
authors.lastname: Shi
authors.forename: Frauke
authors.initials: F
authors.lastname: Rininsland
authors.forename: Stuart
authors.initials: S
authors.lastname: Kushon
authors.forename: Wensheng
authors.initials: W
authors.lastname: Xia
authors.forename: Kevin
authors.initials: K
authors.lastname: Ley
authors.forename: Komandoor
authors.initials: K
authors.lastname: Achyuthan
authors.forename: Duncan
authors.initials: D
authors.lastname: McBranch
authors.forename: David
authors.initials: D
authors.lastname: Whitten
mesh_terms: D016282:Aspartic Acid Endopeptidases / Q000032:analysis*; D020169:Caspases / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid; D003546:Cysteine Endopeptidases / Q000032:analysis*; D004765:Enteropeptidase / Q000032:analysis*; D005453:Fluorescence; D011108:Polymers / Q000378:metabolism*
chemical_list: D011108:Polymers; D004765:Enteropeptidase; D020169:Caspases; D003546:Cysteine Endopeptidases; D016282:Aspartic Acid Endopeptidases