Measurement of Protease Activities Using Fluorogenic Substrates.
Matrix metalloproteinases and the related metalloproteases are implicated in cancer progression. They are endopeptidases that require several defined amino acid residues in both N-terminal and C-terminal sides of the scissile bond. Fluorogenic Förster resonance energy transfer (FRET) substrates that harbor a fluorophore and a quencher on opposite sides of the scissile bond are conveniently used to measure their activities. In this chapter, we describe the principle of FRET substrates and how to use them to measure activities and kinetic parameters of endopeptidases.
['Santamaria|Salvatore|S|', 'Nagase|Hideaki|H|']
D000071121:ADAMTS4 Protein D000595:Amino Acid Sequence D057075:Enzyme Assays D031541:Fluorescence Resonance Energy Transfer D005456:Fluorescent Dyes D007700:Kinetics D009154:Mutation D059748:Proteolysis D011994:Recombinant Proteins D013379:Substrate Specificity
ADAM ADAMTS Enzyme kinetics FRET peptide substrates Inner filter effect MMP