This substrate has a fluorescent Trp residue in subsite Pí2 and a dinitrophenol (Dnp) quenching group at the N-terminus. The quenching of the Trp fluorescence in the intact substrate is relieved upon hydrolysis of the P1-Pí1 bond, giving rise to a continuously recording fluorescence assay. The residues placed in subsites P3-Pí1 and Pí3 is optimized for MMP-3, while Arg is placed in Pí4 to enhance solubility. The substrate hydrolysis rate rivals or exceeds those of the corresponding natural protein substrates or other synthetic peptides. The solubility of this substrate in assay buffer exceeds the Km value for each reaction, allowing accurate determination of the kinetic parameters. Abs/Em = 280/360 nm.
Ref: Finch-Arietta, M. et al. Agents Actions 39, C189 (1993); Netzel-Arnett, S. et al. Anal. Biochem. 195, 86 (1991).
Sequence (One-Letter Code)
Sequence (Three-Letter Code)
Dnp - Pro - Tyr - Ala - Tyr - Trp - Met - Arg - OH