iD SensoLyte® Assay Kits
Furin Activity Assay Kit
Inhibition of furin activity by a peptidyl chloromethylketone measured with the SensoLyte Rh110 Furin Activity Assay Kit (SpectraMax M5 Microplate Reader, Molecular Devices).
- High sensitivity fluorimetric detection of furin activity (0.02 pg/μL of furin)
- For efficient screening of furin inhibitors
- Compatible with HTS for drug discovery applications
- One-step homogeneous mix-and-read format
How Furin Inhibitor Activity Testing Works
- The SensoLyte® Rh110 human Furin Activity Assay Kit employs a fluorescent peptide substrate for the detection of human furin activity. This substrate contains the rhodamine 110 fluorophore (Rh110).
- When Furin cleaves this Rh110 substrate, it results in the release of a bright green fluorescence that can be detected at excitation/emission = 496nm/520 nm. In the presence of an inhibitor, Furin cannot cleave the substrate and no signal is recorded. Long wavelength spectra and the high extinction coefficient of Rh110 provide greater sensitivity and less background interference from other reaction components.
Role of Furin?
- Furin, a proprotein convertase is expressed in all mammalian cell types and processes a variety of precursor molecules into their active forms, including hormones, enzymes, growth factors, receptors, plasma proteins, bacterial toxins, viral envelope proteins, etc.
- Furin has specifically been identified as an anti-viral drug target for HIV or HBV infections.
Why is Furin Important for HIV Assembly?
- Furin is essential for maturation and viral infectivity of HIV. It is one of the proteases which are responsible for the cleavage of the HIV precursor envelope glycoprotein 160 (gp160) into the gp120 and gp41 moieties.
- The gp120 envelope moiety is responsible for virion-specific binding to the cell surface receptor CD4, while gp41 is the transmembrane component that mediates virus-to-cell membrane fusion, thereby causing viral nucleocapsid to penetrate into the cytosol.
- Drugs/inhibitors that inhibit gp160 processing into gp120 and gp41 have been studied to prevent viral propagation.