520 MMP FRET Substrate 6

Product Name: 520 MMP FRET Substrate 6

Sequence One Letter Code: QXL® 520-PLGMWSRK(5-FAM)-NH2

Sequence Three Letter Code: QXL® 520-Pro-Leu-Gly-Met-Trp-Ser-Arg-Lys(5-FAM)-NH2

Molecular Weight: 1823

Purity: 95%

Form: Lyophilized

Storage Conditions: - 20 °C Protected from light

Research Area: peptide substrate

Conjugation: Conjugated

Conjugation Type: Double dyes

Code Nacres: NA.26

Application: 520 MMP FRET Substrate 6 is a fluorogenic peptide substrate developed for selective analysis of matrix metalloproteinase activity. It is efficiently cleaved by MMP-2 and MMP-13, enabling targeted assessment of these enzymes in extracellular matrix and tissue remodeling research. Enzymatic hydrolysis disrupts FRET quenching, producing fluorescence measurable at Ex/Em 494/521 nm. The substrate supports kinetic studies, inhibitor screening, and functional characterization of MMP-2– and MMP-13–mediated proteolytic pathways.

Current Research: 520 MMP FRET Substrate 6 is a fluorogenic peptide substrate engineered for selective and sensitive detection of matrix metalloproteinase (MMP) activity, with preferential responsiveness to MMP-2 (gelatinase A) and MMP-13 (collagenase-3). The substrate incorporates a fluorescence resonance energy transfer (FRET) donor–quencher pair that maintains low background fluorescence in its intact state. Upon proteolytic cleavage, quenching is disrupted, resulting in a measurable fluorescence signal at approximately Ex 494 nm / Em 521 nm. This design supports real-time monitoring of protease activity in biochemical and complex biological systems. Biological Context MMP-2 and MMP-13 are zinc-dependent endopeptidases that play central roles in extracellular matrix (ECM) remodeling: MMP-2 degrades gelatin, type IV collagen, and basement membrane components, contributing to angiogenesis and tumor invasion. MMP-13 exhibits high collagenolytic activity and is implicated in cartilage degradation, osteoarthritis, and tumor-associated matrix remodeling. Selective detection of these enzymes is critical in studies of tissue remodeling, fibrosis, cancer progression, and inflammatory pathology. Mechanism of Detection The substrate contains: An MMP recognition sequence optimized for MMP-2 and MMP-13 cleavage A fluorophore emitting at ~521 nm A proximal quencher suppressing fluorescence via FRET In the intact molecule, energy transfer between donor and quencher prevents fluorescence emission. Upon enzymatic hydrolysis: The peptide bond at the target site is cleaved. Donor and quencher are spatially separated. Fluorescence intensity increases proportionally to enzymatic activity. This enables continuous kinetic measurements with high sensitivity and minimal background interference. Research Applications 1. Targeted MMP-2 and MMP-13 Profiling The substrate is particularly suited for assessing proteolytic activity in systems where gelatinase and collagenase functions are of interest, including cancer invasion and cartilage degeneration models. 2. Extracellular Matrix Remodeling Studies It supports functional analysis of matrix degradation during angiogenesis, wound healing, and fibrotic progression. 3. Inhibitor Screening The robust fluorescence readout is compatible with medium- to high-throughput screening of selective MMP inhibitors. Reduced fluorescence indicates effective enzymatic inhibition. 4. Comparative Protease Analysis The substrate can be used alongside other MMP-selective substrates to distinguish relative contributions of MMP-2 and MMP-13 in mixed protease environments. Advantages Preferential sensitivity to MMP-2 and MMP-13 High signal-to-background ratio due to FRET design Real-time kinetic monitoring capability Suitable for purified enzymes and complex biological samples Compatible with standard fluorescence instrumentation Experimental Considerations Assays require appropriate buffer conditions, including zinc and calcium ions essential for MMP catalytic function. Activation of latent pro-MMPs (e.g., with APMA) may be necessary prior to analysis. Inclusion of selective MMP inhibitors can confirm specificity in mixed protease systems.