Product Details

Purity

≥ 95% as determined by SDS-PAGE.

Endotoxin

Less than 1.0 EU/ug as determined by LAL method.

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 >400 pmol/min/µg.

Target Names

LGMN

Uniprot No.

Q99538

Alternative Names

Legumain; EC 3.4.22.34; Asparaginyl endopeptidase; AEP; Protease, cysteine 1

Species

Homo sapiens (Human)

Source

Mammalian cell

Expression Region

18-433aa

Target Protein Sequence

VPIDDPEDGGKHWVVIVAGSNGWYNYRHQADACHAYQIIHRNGIPDEQIVVMMYDDIAYSEDNPTPGIVINRPNGTDVYQGVPKDYTGEDVTPQNFLAVLRGDAEAVKGIGSGKVLKSGPQDHVFIYFTDHGSTGILVFPNEDLHVKDLNETIHYMYKHKMYRKMVFYIEACESGSMMNHLPDNINVYATTAANPRESSYACYYDEKRSTYLGDWYSVNWMEDSDVEDLTKETLHKQYHLVKSHTNTSHVMQYGNKTISTMKVMQFQGMKRKASSPVPLPPVTHLDLTPSPDVPLTIMKRKLMNTNDLEESRQLTEEIQRHLDARHLIEKSVRKIVSLLAASEAEVEQLLSERAPLTGHSCYPEALLHFRTHCFNWHSPTYEYALRHLYVLVNLCEKPYPLHRIKLSMDHVCLGHY

Mol. Weight

49.0 kDa

Protein Length

Full Length

Tag Info

C-terminal 10xHis-tagged

Form

Lyophilized powder
Note: We will preferentially ship the format that we have in stock, however, if you have any special requirement for the format, please remark your requirement when placing the order, we will prepare according to your demand.

Buffer

Lyophilized from a 0.2 μm sterile filtered 20 mM Tris-HCl, 0.5 M NaCl, 6% Trehalose, pH 8.0

Reconstitution

We recommend that this vial be briefly centrifuged prior to opening to bring the contents to the bottom. Please reconstitute protein in deionized sterile water to a concentration of 0.1-1.0 mg/mL.We recommend to add 5-50% of glycerol (final concentration) and aliquot for long-term storage at -20°C/-80°C. Our default final concentration of glycerol is 50%. Customers could use it as reference.

Troubleshooting and FAQs

Protein FAQs

Storage Condition

Store at -20°C/-80°C upon receipt, aliquoting is necessary for mutiple use. Avoid repeated freeze-thaw cycles.

Shelf Life

The shelf life is related to many factors, storage state, buffer ingredients, storage temperature and the stability of the protein itself.
Generally, the shelf life of liquid form is 6 months at -20°C/-80°C. The shelf life of lyophilized form is 12 months at -20°C/-80°C.

Lead Time

3-7 business days

Notes

Repeated freezing and thawing is not recommended. Store working aliquots at 4°C for up to one week.

Datasheet & COA

Please contact us to get it.

Description

Legumain is a lysosomal cysteine protease with strict specificity for asparagine residues, making it a critical target in cancer biology where its aberrant expression drives tumor invasion and antigen processing. This full-length construct (aa 18–433) demonstrates robust catalytic function with a specific activity exceeding 400 pmol/min/µg against the fluorogenic substrate Z-AAN-AMC, providing a quantitative benchmark for kinetic parameter analysis including Km, Vmax, and kcat/Km determination. The C-terminal His tag preserves access to the active site, supporting use in inhibitor screening campaigns, IC50 determination, and drug candidate evaluation where unobstructed substrate binding is essential. Purity greater than 95% by SDS-PAGE and endotoxin levels below 1.0 EU/µg satisfy the stringent criteria typical for enzymatic assays requiring accurate specific activity measurements and for serving as a positive control in enzyme-linked detection systems.

Target Background

Function

Has a strict specificity for hydrolysis of asparaginyl bonds. Can also cleave aspartyl bonds slowly, especially under acidic conditions. Required for normal lysosomal protein degradation in renal proximal tubules. Required for normal degradation of internalized EGFR. Plays a role in the regulation of cell proliferation via its role in EGFR degradation. May be involved in the processing of proteins for MHC class II antigen presentation in the lysosomal/endosomal system.

Gene References into Functions

Studies identified Legumain (LGMN) as a new target which is highly expressed in the tumor microenvironment and in tumor cells. [review] PMID: 27993111
our results suggest that M2 tumour-associated macrophages affect degradation of the extracellular matrix and angiogenesis via overexpression of legumain, and therefore play an active role in the progression of DLBCL. PMID: 27464733
Legumain regulates oxLDL-induced macrophage apoptosis by enhancing the autophagy pathway. PMID: 29414692
MiRNA-3978 regulates peritoneal gastric cancer metastasis by targeting legumain expression, promoting cell migration/neoplasm invasiveness. PMID: 27793040
These results showed that Asparaginyl endopeptidase could promote invasion and metastasis by modulating epithelial-to-mesenchymal transition. PMID: 27102302
Our data suggest that altered proteolytic activity of legumain in the bone microenvironment contributes to decreased bone mass in postmenopausal osteoporosis. PMID: 28162997
Legumain is increased in both plasma and plaques of patients with carotid stenosis and might be a new and early biomarker of atherosclerosis. PMID: 27940038
Data suggest that melanoma cells internalize/absorb cystatin C from culture media, leading to increased intracellular cystatin C levels; cystatin E/M is internalized as well but at modest rate due to down-regulation of cell migration; however, the effect of intracellular cystatin E/M on down-regulation of legumain activity is pronounced. PMID: 28630039
AEP is activated and cleaves human alpha-synuclein at N103 in an age-dependent manner. PMID: 28671665
AEP promotes activation of the PI3K-AKT signaling pathway in prostate cancer cells. PMID: 27590439
upregulation of legumain is associated with malignant behavior of uveal melanoma. PMID: 26846877
Studies indicate that legumain, usually in lysosomes, is also found extracellularly and even translocates to the cytosol and the nucleus. PMID: 26403494
legumain might play an important role in cervical cancer cell migration and invasion. PMID: 26802645
Legumain appears to be involved in tumor development and deterioration. PMID: 26607955
AEP acts as a delta-secretase, cleaving APP at N373 and N585 residues, selectively influencing the amyloidogenic fragmentation of APP. AEP contributes to the age-dependent pathogenic mechanisms in Alzheimer disease. PMID: 26549211
High legumain expression is associated with breast cancer. PMID: 25605174
This unique feature was confirmed by the crystal structure of AEPpH4.5 (AEP was matured at pH 4.5 and crystallized at pH 8.5), in which the broken peptide bonds were religated and the structure was transformed back to its proenzyme form. PMID: 24407422
High legumain activity is associated with breast cancer. PMID: 24742492
AEP acts as a crucial mediator of tau-related clinical and neuropathological changes. PMID: 25326800
Identified an alternative oncogenic pathway for TRAF6 that uses AEP as its substrate. AEP and TRAF6 protein levels may have prognostic implications in breast cancer patients. Thus, AEP may serve as a biomarker as well as new therapeutic target PMID: 24610907
the relationship among Legumain expression, clinicopathologic, biological variables and patient prognosis in gastric carcinoma PMID: 24023813
HCT116 cells were transfected with p53 siRNA and the effect of knockdown of p53 expression on legumain expression was examined. The results showed that expression levels of both legumain mRNA and protein were decreased in the siRNA-treated cells. PMID: 23942113
Prostate cancer with a vesicular staining pattern of legumain had the potential of being highly invasive and aggressive in patients treated with radical prostatectomy. This suggests that legumain might contribute to the invasiveness and aggressiveness. PMID: 23124822
The multibranched and context-dependent activation process of legumain illustrates how proteases can act not only as signal transducers but as decision makers. PMID: 23776206
Involvement of brain acidosis in the etiopathogenesis of Alzheimer disease, and the asparaginyl endopeptidase-I2(PP2A)-protein phosphatase 2A-Tau hyperphosphorylation pathway. PMID: 23640887
glycosaminoglycans accelerated the autocatalytic activation of prolegumain PMID: 23160071
In the HCT116 and SW620 cell lines nuclear Data indicate that legumain was found to make up approximately 13% and 17% of the total legumain, respectively. PMID: 23326369
Data show a regulatory role of cystatin E/M in controlling both intra- and extracellular legumain activity. PMID: 22902879
TDP-43 is cleaved by AEP in brain. PMID: 22718532
Increased legumain expression was validated by real-time PCR and Western blots, correlated positively with an increased malignancy of ovarian tumors. PMID: 22441772
Accepting asparagines and, to lesser extent, aspartic acid in P1, super-activated legumain exhibits a marked pH dependence that is governed by the P1 residue of its substrate and conformationally stabilizing factors such as temperature or ligands PMID: 22232165
GARP is a key receptor controlling FOXP3 in T(reg) cells following T-cell activation in a positive feedback loop assisted by LGALS3 and LGMN PMID: 19453521
The Lgmn can serve as novel target in diabetes mellitus genetic therapy. PMID: 20536387
the level of cystatin E/M regulates legumain activity and hence the invasive potential of human melanoma cells PMID: 20074384
Legumain is expressed in both murine and human atherosclerotic lesions, may play a functional role in atherogenesis PMID: 18377911
human monocyte-derived dendritic cells harbor inactive proforms of AEP (legumain) that become activated upon maturation of dendritic cells with lipopolysaccharide PMID: 12860980
Freshly isolated human B lymphocytes lack significant asparagine-specific endoprotease (AEP/legumain) activity; cleavage by AEP is dispensable for proteolytic processing of myelin basic protein in this type of cell. PMID: 15100291
Increased legumain expression is associated with primary colorectal cancer PMID: 15788679
AEP may be a proteinases activated by acidosis triggering neuronal injury during neuroexcitotoxicity or ischemia. PMID: 18374643

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Subcellular Location

Lysosome.

Protein Families

Peptidase C13 family

Tissue Specificity

Ubiquitous. Particularly abundant in kidney, heart and placenta.

Database Links

HGNC: 9472

OMIM: 602620

KEGG: hsa:5641

STRING: 9606.ENSP00000334052

UniGene: Hs.18069

Code	CSB-MP012903HU(A4)
Abbreviation	Recombinant Human LGMN protein (Active)
MSDS	MSDS Datasheet
Size	$138
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Image	(Tris-Glycine gel) Discontinuous SDS-PAGE (reduced) with 5% enrichment gel and 15% separation gel. 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 >400 pmol/min/µg. Biological Activity Assay
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Recombinant Human Legumain (LGMN) (Active)

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