Recombinant Human MTAP His-tag Protein, CF

R&D Systems | Catalog # 10379-MT

R&D Systems
100% Guarantee

Key Product Details

  • R&D Systems E. coli-derived Recombinant Human MTAP His-tag Protein (10379-MT)
  • 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

Source

E. coli

Accession Number

Q13126-1

Applications

Enzyme Activity
View all MTAP Proteins and Enzymes »

Product Specifications

Source

E. coli-derived human MTAP protein
Ala2-His283
with an N-terminal Met and 6-His tag

Purity

>95%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

<0.10 EU per 1 μg of the protein by the LAL method.

N-terminal Sequence Analysis

Met

Predicted Molecular Mass

32 kDa

SDS-PAGE

30-32 kDa under reducing conditions

Activity

Measured by its ability to produce adenine through the conversion of 5'methylthioadenine to 5'methylthioribose-1-phosphate.
The specific activity is >650 pmol/min/μg, as measured under the described conditions.

Scientific Data Images for Recombinant Human MTAP His-tag Protein, CF

Recombinant Human MTAP His-tag Protein Enzyme Activity

Recombinant Human MTAP His-tag Protein Enzyme Activity

Recombinant Human MTAP His-tag (Catalog # 10379-MT) is measured by its ability to produce adenine through the conversion of 5'methylthioadenine to 5'methylthioribose-1 phosphate.
Recombinant Human MTAP His-tag Protein SDS-PAGE

Recombinant Human MTAP His-tag Protein SDS-PAGE

2 μg/lane of Recombinant Human MTAP His-tag (Catalog # 10379-MT) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 30-32 kDa under reducing conditions.

Formulation, Preparation, and Storage

10379-MT
Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl and TCEP.
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.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Background: MTAP

Methyl-thioadenosine phosphorylase/MTAP is part of the PNP/MTAP phosphorylase family and catalyzes the reversible phosphorylation of S-methyl-5'-thioadenosine (MTA), a major byproduct of polyamine synthesis essential for cell growth and proliferation. MTAP also produces most of the free adenine generated in human cells through a salvage pathway and thus couples the purine salvage pathway with polyamine biosynthesis. MTAP forms an active trimer where each identical 32 kDa monomer contains a separate active site (1). Each active site contains three distinct regions required for base-, methylthioribose-, and sulfate/phosphate-binding (1). MTAP is cytosolic and abundantly expressed in normal cells and tissues (2). In contrast, deficient MTAP expression is observed in many types of tumors including lung, bladder, pancreatic, and endometrial cancer (3) due to hyper-methylation gene suppression (4) or gene deletion (3, 5, 6). MTA accumulation leads to an immunosuppressive tumor microenvironment and apoptotic resistance (7-9) and MTAP directly regulates the level of MTA present. MTAP has been reported as a tumor suppressor (6,10) that may also act in a manner that is independent of enzymatic activity (11) through signaling pathways such as the insulin-like growth factor-1 receptor pathway (12). Potential therapeutic strategies to exploit MTAP deficiency in tumors (13,14) or inhibit MTAP in tumors that express MTAP, such as prostate cancer, are under investigation (15).

References

  1. Appleby, T.C. et al. (1999) Structure 7:629.
  2. Garbers, D.L. (1978) Biochim. Biophys. Acta. 523:82.
  3. Li, Y. et al. (2019) J. Cancer 10:927.
  4. Hellerbrand, C. et al. (2006) Carcinogenesis 27:64.
  5. Cairns, P. et al. (1995) Nat. Genet. 11:210.
  6. Christopher, S.A. (2002) Cancer Res. 62:6639.
  7. Kirovski, G. et al. (2011) Am. J. Pathol. 178:1145.
  8. Czech, B. et al. (2013) PLoS One 8:e80703.
  9. Henrich, F.C. et al. (2016) Oncoimmunology 5:e1184802.
  10. Kadariya, Y. et al. (2009) Cancer Res. 69:5961.
  11. Tang, B. et al. (2014) G3 (Bethesda) 5:35.
  12. Xu, J. et al. (2019) Signal Transduct. Target Ther. 4:2.
  13. Bertino, J.R. et al. (2011) Cancer Biol. Ther. 11:627.
  14. Tang, B. et al. (2018) Cancer Res. 78:4386.
  15. Bistulfi, G. et al. (2016) Oncotarget 7:14380.

Long Name

5'-Methylthioadenosine Phosphorylase

Alternate Names

BDMF, C86fus, DMSFH, DMSMFH, HEL-249, LGMBF, MTAPase

Entrez Gene IDs

4507 (Human); 66902 (Mouse)

Gene Symbol

MTAP

UniProt

Q13126-1

Additional MTAP Products

Product Documents for Recombinant Human MTAP His-tag Protein, CF

Certificate of Analysis

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

Note: Certificate of Analysis not available for kit components.

Download SDS

Product Specific Notices for Recombinant Human MTAP His-tag Protein, CF

For research use only

Related Research Areas

Sulfation Regulators

Customer Reviews for Recombinant Human MTAP His-tag Protein, CF

There are currently no reviews for this product. Be the first to review Recombinant Human MTAP His-tag Protein, CF and earn rewards!

Have you used Recombinant Human MTAP His-tag Protein, CF?

Submit a review and receive an Amazon gift card!

$25/€18/£15/$25CAN/¥2500 Yen for a review with an image

$10/€7/£6/$10CAN/¥1110 Yen for a review without an image

Submit a review
Amazon Gift Card

Protocols

View specific protocols for Recombinant Human MTAP His-tag Protein, CF (10379-MT):

Materials
  • Assay Buffer: 50 mM Potassium Phosphate, 1 mM DTT, pH 7.4
  • Recombinant Human Methylthioadenosine Phosphorylase (rhMTAP) (Catalog # 10379-MT)
  • 5'-Deoxy-5'-Methylthioadenosine (MTA) (Cayman Chemical, Catalog # 15593), 30 mM stock in DMSO
  • Xanthine Oxidase (Sigma, Catalog # X2252), 200 U/mL stock in 10 mM Potassium Phosphate, pH 7.4
  • UV Plate (Costar, Catalog # 3635)
  • Plate Reader (Model: SpectraMax Plus by Molecular Devices) or equivalent
  1. Dilute rhMTAP to 2 µg/mL in Assay Buffer.
  2. Prepare Substrate Mixture containing 600 µM MTA and 6 U/mL Xanthine Oxidase in Assay Buffer.
  3. Load into a plate 50 µL of 2 µg/mL rhMTAP, and start the reaction by adding 50 µL of Substrate Mixture. Include a Substrate Blank containing 50 µL of Assay Buffer and 50 µL of Substrate Mixture.
  4. Seal the plate and incubate at room temperature for 15 minutes.
  5. Read plate at 305 nm (absorbance) in kinetic mode for 5 minutes.
  6. Calculate specific activity:

     Specific Activity (pmol/min/µg) =

Adjusted Vmax* (OD/min) x well volume (L) x 1012 pmol/mol

ext. coeff** (M-1cm-1) x path corr.*** (cm) x amount of enzyme (µg)

*Adjusted for Substrate Blank
**Using extinction coefficient 15500 M-1cm-1
***Using the path correction 0.32 cm

Note:  the output of many spectrophotometers is in mOD

Per Well:
  • rhMTAP: 0.1 µg
  • MTA: 300 µM
  • Xanthine Oxidase: 3 U/mL




















FAQs

No product specific FAQs exist for this product.

View all FAQs for Proteins and Enzymes