Quenched fluorescent substrate-based peptidase assays.
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.
Lew
Rebecca A
RA
Department of Biochemistry and Molecular Biology, Monash University, Clayton, Victoria, Australia.
Tochon-Danguy
Nathalie
N
Hamilton
Catherine A
CA
Stewart
Karen M
KM
Aguilar
Marie-Isabel
MI
Smith
A Ian
AI
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
D019332:Endothelin-1; D005456:Fluorescent Dyes; D010455:Peptides; D010447:Peptide Hydrolases; D016282:Aspartic Acid Endopeptidases; D008666:Metalloendopeptidases; D000072396:Endothelin-Converting Enzymes; D001920:Bradykinin