Recombinant A. Muciniphila M60 domain-containing peptidase
Recombinant A. Muciniphila M60 domain-containing peptidase Summary
- R&D Systems E. coli-derived Recombinant A. Muciniphila M60 domain-containing peptidase (11479-M6)
- Quality control testing to verify active proteins with lot specific assays by in-house scientists
- All R&D Systems proteins are covered with a 100% guarantee
Product Specifications
Ala21-Glu506 with an N-terminal Met and C-terminal 6-His tag
Analysis
Product Datasheets
Carrier Free
CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.
In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.
11479-M6
| Formulation | Supplied as a 0.2 μm filtered solution in Tris and NaCl. |
| Shipping | The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below. |
| Stability & Storage: | Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
|
Assay Procedure
- Assay Buffer: 50 mM MES, pH 6.5
- Recombinant A. Muciniphila M60 domain-containing peptidase His-tag (rAm M60 peptidase) (Catalog # 11479-M6)
- Substrate: Recombinant Human CA125/MUC16 Protein (rhMUC16) (Catalog # 5609-MU)
- 4-20% SDS-PAGE Gel Gel loading dye
- Gel Imager/Densitometer
- Dilute rAm M60 peptidase to 25 µg/mL with Assay Buffer.
- Dilute rhMUC16 to 100 µg/mL with Assay Buffer.
- Combine 10 µL of rAm M60 peptidase and 10 µL of rhMUC16 in a tube.
- Create a negative control by adding 10 µL of rhMUC16 and 10 µL of Assay Buffer.
- Incubate reactions and controls at 37 °C for 24 hours.
- Add gel loading dye to all tubes.
- Load the total reaction volume on a 4-20% SDS-PAGE gel and run down 80% of the gel, minimally.
- Stain the gel and acquire image.
- Use densitometry to calculate the percent digestion of rhMUC16 by rAm M60 peptidase.
- rAm M60 peptidase: 0.25 μg
- rhMUC16: 1 μg
Scientific Data
View Larger
Recombinant A. Muciniphila M60 domain-containing peptidase His-tag Protein (Cat # 11479-M6) cleaves glycoproteins N-terminally to glycosylated Ser/Thr residues that are near but not next to the cleavage residue. Specificity requires two adjacent truncated O-glycans and M60 peptidase cleaves between the two residues. * represents the residue for which M60 peptidase has recognition. Sialyated glycan substrates in diagram are less preferred.
View Larger
Recombinant A. Muciniphila M60 domain-containing peptidase His-tag Protein (Cat # 11479-M6) is measured by its ability to digest Recombinant Human CA125/MUC16 Protein, CF (5609-MU) and detected via SDS-PAGE gel.
View Larger
2 μg/lane of Recombinant A. Muciniphila M60 domain-containing peptidase His-tag Protein (Catalog # 11479-M6) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 52-57 kDa, under reducing conditions.
Reconstitution Calculator
Background: muciniphila M60 domain-containing
Recombinant A. Muciniphila M60 domain-containing peptidase (Am M60 peptidase), also referred to as AM0627, is a zinc-dependent mucin-targeting protease from Akkermansia muciniphila, a gut symbiont found within the human and mouse gut mucus layer. Am M60 peptidase is classified as an enzyme within the large M60-like family of metalloproteases containing gluzincin motifs from organisms known to inhabit mucosal host environments (1,2). Am M60 peptidase is a secreted, monomeric protein with a signal sequence, an immunoglobulin-like fold domain, and catalytic domain that contains the zinc-binding site within the gluzincin motif (2,3). Am M60 peptidase is capable of cleaving glycopeptides between adjacent O-glycosylated threonine or serine, with a preference towards desialylated substrates (4). Glycopeptides with adjacent non-physiological truncated O-glycan structure that promote tumorigenesis and metastasis, known as tumor-associated carbohydrate antigens, serve as optimal substrates for Am M60 peptidase (3,5). A. muciniphilia relies on host-derived mucin as an energy source and leads to production of short chain fatty acids (SCFA) via mucin-degradation byproducts. SCFA regulate inflammation and provide hosts with health benefits for diseases such as inflammatory bowel disease, obesity, autism, and cancer (2, 6-9). Mucin-degrading enzymes such as Am M60 peptidase play a prominent role in host mucin degradation and consequently Am M60 peptidase may be of interest as a therapeutic target, a diagnostic tool to detect and monitor disease progression, and/or useful as a tool to further study mucins and O-glycoproteins (2-4, 10).
- Rawlings, N.D. et al. (2014) Nucl. Acids Res. 42: D503.
- Shon, D.J. et. al. (2022) J. Biol. Chem. 298: 101917.
- Taleb, V. et. al. (2022) Nat. Commun. 13: 4324.
- Shon, D.J. et. al. (2020) Proc. Natl. Acad USA 117: 21299.
- Kudelka, M.R. et. al. (2015) Adv. Cancer Res. 126:53.
- Yamada, T. et al. (2019) EbioMedicine 48: 513.
- Hansson, G.C. et. al. (2020) Annu. Rev. Biochem. 89: 769.
- Pruss, K.M. et. al. (2021) ISME J. 15: 577.
- Molaaghaee-Rouzbahani, S. et. al. (2023) Sci. Rep. 13:3237.
- Labourel, A. et. al. (2023) Essays Biochem. 67:331.
FAQs
No product specific FAQs exist for this product, however you may
View all Proteins and Enzyme FAQsReviews for Recombinant A. Muciniphila M60 domain-containing peptidase
There are currently no reviews for this product. Be the first to review Recombinant A. Muciniphila M60 domain-containing peptidase and earn rewards!
Have you used Recombinant A. Muciniphila M60 domain-containing peptidase?
Submit a review and receive an Amazon gift card.
$25/€18/£15/$25CAN/¥75 Yuan/¥2500 Yen for a review with an image
$10/€7/£6/$10 CAD/¥70 Yuan/¥1110 Yen for a review without an image
