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Matrix metalloproteinase-3 (MMP-3) fluorometric drug discovery kit, RED
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The MMP-3 Fluorometric (also known as fluorimetric) Drug Discovery Kit, RED is a complete assay system designed to screen MMP-3 inhibitors using a quenched fluorogenic substrate OMNIMMP® RED: TQ3-GABA-Pro-Cha-Abu-Smc-His-Ala-Dab(6-TAMRA)-Ala-Lys-NH2 [TQ3=quencher; GABA=4-aminobutyric acid; Cha=L-cyclohexylalanine; Abu=2-aminobutyric acid; Smc=S-methyl-L-cysteine; Dab=2,4-diaminobutyric acid; 6-TAMRA=6-tetramethylrhodamine]. TAMRA fluorescence is thoroughly quenched by the TQ3 group until cleavage by MMPs separates the two moieties. The assays are performed in a convenient 96-well microplate format. The kit is useful to screen inhibitors of MMP-3, a potential therapeutic target. The compound NNGH is also included as a prototypic control inhibitor.Matrix metalloproteinase-3 (MMP-3, stromelysin-1, transin-1) is a member of the MMP family of extracellular proteases. These enzymes play a role in many normal and disease states by virtue of their broad substrate specificities. Targets of MMP-3 include collagens, fibronectin, and laminin, plasminogen, HB-EGF, E-cadherin, and other MMPs. MMP-3 is secreted as a 55-59kDa glycosylated proenzyme (measured by SDS-PAGE), and activated by cleavage to forms of 21-48kDa. It is unique from other MMPs in that its pH optimum is 5.9, rather than around 7.0.
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| Alternative Name Stromelysin-1, Transin-1 | Application Fluorescent detection, HTS | Application Notes Designed to screen MMP-3 inhibitors using a quenched fluorogenic peptide. | Contents 1 vial MMP-3 enzyme1 vial substrate (OMNIMMP® RED)1 vial 6′-TAMRA calibration standard1 vial control inhibitor (NNGH)1 bottle (20 ml) assay buffer1 black 96-well microplateInstructions | Technical Info / Product Notes NCBI Reference Sequence: NM_002422The OMNIMMP® RED substrate offers key advantages over other MMP substrates.{{1640282-42-3} web|{1640282-42-3} Protocol|{1640282-42-3} References|{1640282-42-3} manufacturer} Emission at the red end of the spectrum (576 nm after excitation at 545 nm) avoids the interference at lower wavelengths often exhibited by screening compounds, and by substances commonly found in biological samples and tissue culture medium.{{1599440-33-1} web|{1599440-33-1} Biological Activity|{1599440-33-1} Description|{1599440-33-1} supplier} MMP substrate peptides display poor aqueous solubility, often with Kms near their limits of solubility, making enzyme and inhibitor kinetics difficult.PMID:31078606 MMP Kms for OMNIMMP® RED substrate are below its solubility limit.OMNIMMP® RED is avidly cleaved by MMPs, with kcat/Kms in the range of 104-106 M-1sec-1. The ultra-strong fluorescence of OMNIMMP® RED allows for substrate concentrations much lower than the Km, a condition generally desirable in inhibitor screening assays. | UniProt ID P08254
