title: Quenched fluorescent substrate-based peptidase assays.
abstract: The use of specific quenched fluorescent substrates (QFS) provides a rapid and sensitive method to measure peptidase activity, and is readily adaptable to high-throughput screening of potential peptidase inhibitors. In this chapter, we discuss general considerations for the development of QFS assays, and describe in detail an assay protocol for the mammalian metallopeptidase, endothelin-converting enzyme.
authors.affiliation: Department of Biochemistry and Molecular Biology, Monash University, Clayton, Victoria, Australia.
authors.forename: Rebecca A
authors.initials: RA
authors.lastname: Lew
authors.forename: Nathalie
authors.initials: N
authors.lastname: Tochon-Danguy
authors.forename: Catherine A
authors.initials: CA
authors.lastname: Hamilton
authors.forename: Karen M
authors.initials: KM
authors.lastname: Stewart
authors.forename: Marie-Isabel
authors.initials: MI
authors.lastname: Aguilar
authors.forename: A Ian
authors.initials: AI
authors.lastname: Smith
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D016282:Aspartic Acid Endopeptidases / Q000032:analysis / Q000378:metabolism; D001920:Bradykinin / Q000737:chemistry; D019332:Endothelin-1 / Q000737:chemistry; D000072396:Endothelin-Converting Enzymes; D005456:Fluorescent Dyes / Q000737:chemistry; D008666:Metalloendopeptidases / Q000032:analysis / Q000378:metabolism; D008967:Molecular Biology / Q000379:methods; D008969:Molecular Sequence Data; D010447:Peptide Hydrolases / Q000032:analysis* / Q000378:metabolism; D010455:Peptides / Q000737:chemistry; D013379:Substrate Specificity
chemical_list: D019332:Endothelin-1; D005456:Fluorescent Dyes; D010455:Peptides; D010447:Peptide Hydrolases; D016282:Aspartic Acid Endopeptidases; D008666:Metalloendopeptidases; D000072396:Endothelin-Converting Enzymes; D001920:Bradykinin