Furin

Description

Furin is a ubiquitous subtilisin-like proprotein convertase. It is the major processing enzyme of the secretory pathway and is localized in the trans-golgi network (1,2). Substrates of Furin include blood clotting factors, serum proteins and growth factor receptors such as the insulin-like growth factor receptor (3). The minimal cleavage site is Arg-X-X-Arg'. However, the enzyme prefers the site Arg-X-(Lys/Arg)-Arg'. An additional arginine at the P6 position appears to enhance cleavage (4). Furin is inhibited by EGTA, α1- Antitrypsin Portland (5) and polyarginine compounds (6).

Product Source

Isolated from Spodoptera frugiperda (Sf9) cells infected with recombinant baculovirus carrying truncated human furin (kindly provided by R. Fuller) (3).

Properties and Usage

Unit Definition

1 unit is defined as the amount of enzyme required to cleave 25 µg of a MBP-FN-paramyosin-ΔSal substrate to 95% completion in 6 hours at 25°C in a total reaction volume of 25 µl.

Storage Temperature

-20°C

Storage Conditions

10 mM MES
1 mM CaCl2
50% Glycerol
pH 7.0 @ 25°C

Molecular Weight

Theoretical: 52.7 kDa

Unit Assay Conditions

Two fold dilutions of Furin are incubated with 25 µg MBP-FN-paramyosin-ΔSal substrate in 20 mM HEPES, 0.1% Triton X-100, 0.2 mM CaCl2. 0.2mM β-mercaptoenthanol (pH 7.5 @ 25°C) in a 25 µl reaction. The reaction mix is incubated at 25°C for 6 hours. Separation of reaction products are visualized by SDS-PAGE.

Notes

  1. Both Furin and Onchocerca volvulus Blisterase will cleave peptide substrates with the sequence, Arg-X- (Lys/Arg)-Arg. However, the ability of either enzyme to cleave a particular protein substrate depends on its tertiary structure as well as on the amino acids immediately surrounding the cleavage site (7).

References

  1. van den Ouweland, A.M.W. et al. (1990). Nucl. Acids Res. 18, 664.
  2. Steiner, D.F. (1998). Curr. Opin. Chem. Biol. . 2
  3. Bravo, D.A. et al. (1994). J. Biol. Chem. . 269, 25830-25837.
  4. Krysan, D.J. et al. (1999). J. Biol. Chem.. 274, 23229-23234.
  5. Jean, F. et al. (1998). Proc. Natl. Acad. Sci. USA. 95, 7293-7298.
  6. Cameron, A. et al. (2000). J. Biol. chem. . 275, 36741-36749.
  7. Poole, C.B. et al. (2003). J. Biol. Chem. . 278, 36183-36190.

Datacards

The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.

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Specifications

The Specification sheet is a document that includes the storage temperature, shelf life and the specifications designated for the product. The following file naming structure is used to name these document files: [Product Number]_[Size]_[Version]

Safety Data Sheet

The following is a list of Safety Data Sheet (SDS) that apply to this product to help you use it safely.

Datacards

The Product Summary Sheet, or Data Card, includes details for how to use the product, as well as details of its formulation and quality controls. The following file naming structure is used to name the majority of these document files: [Catalog Number]Datasheet-Lot[Lot Number]. For those product lots not listed below, please contact NEB at info@neb.com or fill out the Technical Support Form for appropriate document.