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Recombinant Mouse Legumain/Asparaginyl Endopeptidase, CF

Bio-Techne includes R&D Systems | Catalog # 2058-CY

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2058-CY-010

Key Product Details

Source

NS0

Accession #

Structure / Form

Pro form

Conjugate

Unconjugated

Applications

Enzyme Activity

Product Specifications

Source

Mouse myeloma cell line, NS0-derived mouse Legumain/Asparaginyl Endopeptidase protein
Val18-Tyr435, with an N-terminal 7-His tag

Purity

>95%, by SDS-PAGE under reducing conditions and visualized by silver stain.

Endotoxin Level

<1.0 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

His

Predicted Molecular Mass

49 kDa

SDS-PAGE

61 kDa, reducing conditions

Activity

Measured by its ability to cleave the fluorogenic peptide substrate, N-carbobenzyloxy-Ala-Ala-Asn-7-amido-4-methylcoumarin (Z-AAN-AMC).
The specific activity is >350 pmol/min/µg, as measured under the described conditions.

Formulation, Preparation and Storage

2058-CY
Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl and Glycerol.
Shipping The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Background: Legumain/Asparaginyl Endopeptidase

Legumain is a lysosomal cysteine protease found in all mouse tissues examined, but was particularly abundant in kidney and placenta (1). Legumain plays a pivotal role in the endosomal/lysosomal degradation system because the Legumain deficiency causes the accumulation of pro cathepsins B, H and L, another group of lysosomal cysteine proteases (2). Over-expression of Legumain in tumors is significant for invasion/metastasis (3). Also known as asparaginyl endopeptidase, it specifically cleaves peptide bonds with Asn at the P1 position. Nevertheless, it also cleaves peptide bonds with Asp at the P1 position. Auto-activation of pro Legumain involves both types of cleavage, which results in the removal of the pro peptides in both C- and N-termini (4). In addition, Legumain activates pro MMP-2 and processes bacterial antigens for MHC class II presentation and pro thymosin alpha to thymosin alpha1 and thymosin alpha11, two acidic peptides with immunoregulatory properties (5-7). Mouse Legumain is synthesized as a 435 amino acid precursor with a signal peptide (residues 1 to 17). The pro enzyme (residues 18 to 435) was expressed with an N-terminal His tag. The purified pro enzyme can be activated under the conditions as described above. Legumain activity can be inhibited by rmCystatin C and recombinant human cystatins C and E/M (Catalog # 1238-PI, 1196-PI, and 1286-PI). 

References

  1. Chen, J.M. et al. (1998) Biochem. J. 335:111.
  2. Shirahama-Noda, K. et al. (2003) J. Biol. Chem. 278:33194.
  3. Liu, C. et al. (2003) Cancer Res. 63: 2957.
  4. Li D.N. et al. (2003) J. Biol. Chem. 278:38980.
  5. Chen, J.M. et al. (2001) Biol. Chem. 382:777.
  6. Schwarz, G. et al. (2002) Biol. Chem. 383:1813.
  7. Sarndeses, C.S. et al. (2003) J. Biol. Chem. 278:13286.

Alternate Names

AEP, Asparaginyl Endopeptidase, LGMN

Entrez Gene IDs

5641 (Human); 19141 (Mouse)

Gene Symbol

LGMN

UniProt

Additional Legumain/Asparaginyl Endopeptidase Products

Product Documents for Recombinant Mouse Legumain/Asparaginyl Endopeptidase, CF

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Recombinant Mouse Legumain/Asparaginyl Endopeptidase, CF

For research use only

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