Recombinant Human MGAT4A His-tag, CF

R&D Systems | Catalog # 11496-GT

Alpha-1,3-mannosyl-glycoprotein 4-beta-N-acetylglucosaminyltransferase A
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Key Product Details

  • R&D Systems CHO-derived Recombinant Human MGAT4A His-tag (11496-GT)
  • 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

CHO

Accession Number

Q9UM21

Applications

Enzyme Activity
View all MGAT4A Proteins and Enzymes »

Product Specifications

Source

Chinese Hamster Ovary cell line, CHO-derived human MGAT4A protein
Leu93-Asn535, with a C-terminal 6-His tag

Purity

>90%, 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

Leu93

Predicted Molecular Mass

52 kDa

SDS-PAGE

54-60 kDa, under reducing conditions.

Activity

Measured by its ability to transfer N-acetyl-alpha -D-glucosamine from UDP-N-Acetyl-alpha -D-glucosamine to glycan Cy5-Fuc labeled N2f.
Able to convert >85% of the substrate glycan N2f, as measured under the described conditions.

Scientific Data Images for Recombinant Human MGAT4A His-tag, CF

Recombinant Human MGAT4A His-tag Protein Enzyme Activity Diagram.

Recombinant Human MGAT4A His-tag Protein (Catalog # 11496-GT) catalyzes a beta 1-4GlcNAc linkage to the alpha 1-3Man arm of N-glycans.

Recombinant Human MGAT4A His-tag Protein Enzyme Activity.

Recombinant Human MGAT4A His-tag Protein (Catalog # 11496-GT) activity was measured by its ability to transfer N-acetyl-alpha -D-glucosamine from UDP-N-acetyl-alpha -D-glucosamine to glycan Cy5-Fuc labeled N2f (GL304). Following treatment, a mobility shift was observed via SDS-PAGE gel.

Recombinant Human MGAT4A His-tag Protein SDS-PAGE.

2 μg/lane of Recombinant Human MGAT4A His-tag Protein (Catalog # 11496-GT) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 54-60 kDa, under reducing conditions.

Formulation, Preparation, and Storage

11496-GT
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.
  • 6 months from date of receipt, -20 to -70 °C as supplied.
  • 3 months, -20 to -70 °C under sterile conditions after opening.

Background: MGAT4A

Recombinant human Alpha-1,3-mannosyl-glycoprotein 4-beta -N-acetylglucosaminyltransferase A (MGAT4A), also known as GlcNAc-T IVa, is a metal-dependent, golgi single-pass type II membrane protein that contains an N-terminal transmembrane region, a catalytic domain, and a C-terminal carbohydrate binding module that regulates its catalytic activity (1, 2). MGAT4A is one of several human MGAT enzymes involved in the initiation and synthesis of N-glycans, with each enzyme having unique specificity and preferences (3). MGAT4 has two human isozymes that catalyze formation of the beta 1-4 GlcNAc branch in the  alpha 1-3 Mannose arm of N‑glycans (4-6). The two isozymes have different expression profiles in tissues and cancer cell lines, with MGAT4A expression within gastrointestinal tissues such as the pancreas (2, 4, 5). Loss of MGAT4A in mouse models has been shown to induce type 2 diabetes (9,10). Expression and activity of MGAT4A is particularly important during differentiation and oncogenesis (7, 8) including within choriocarcinoma, invasive mole, and placental site trophoblastic tumors (11-13). The discovery of MGAT4A modulators may lead to new therapeutics for the treatment of these diseases (10). MGAT4A may also be useful as a tool for the glycoengineering of complex N-glycans. The activity of MGAT4A was demonstrated using a fluorescent gel shift assay.

References

  1. Oka, N. et al. (2022) Glycobiology. 32:1153.
  2. Nagae, M. et. al. (2022) Commun. Biol. 5:695.
  3. Brockhausen, I. et. al. (1988) Biochemie 70:1521.
  4. Yoshida, A. et. al. (1999) Glycobiology. 9:303.
  5. Oguri, S. et. al. (2006) Glycoconj. J. 23:473.
  6. Osada, N. et. al. (2022) J. Biol. Chem. 298:102400.
  7. Nishikawa, A. et. al. (1990) Biochim. Biophys. Acta. 1035:313.
  8. Nakao, H. et. al. (1990) Biochem. Biophys. Res. Commun. 172:1260.
  9. Ohtsubo, K. et. al. (2005) Cell. 123:1307.
  10. Ohtsubo, K. et. al. (2011) Nat. Med. 17:1067.
  11. Mizuochi, T. et. al. (1983) J. Biol. Chem. 258:14126.
  12. Endo, T. et. al. (1987) Cancer Res. 47:5242.
  13. Niimi, K. et. al. (2012) Br. J. Cancer. 107:1969.

Long Name

alpha-1,3-mannosyl-glycoprotein 4-beta-N-acetylglucosaminyltransferase A

Alternate Names

EC 2.4.1.145, GlcNAc-T IVa, GnT-4a, GNT-IV, GNT-IVA, N-Acetylglucosaminyltransferase IVa

Entrez Gene IDs

11320 (Human); 269181 (Mouse); 367252 (Rat)

Gene Symbol

MGAT4A

UniProt

Q9UM21 (Human)

Additional MGAT4A Products

Product Documents for Recombinant Human MGAT4A His-tag, CF

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Recombinant Human MGAT4A His-tag, CF

For research use only

Related Research Areas

Glycobiology-related Enzymes

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Protocols

View specific protocols for Recombinant Human MGAT4A His-tag, CF (11496-GT):

Materials
  • Assay Buffer: 50 mM Tris, 10 mM MnCl2, pH 7.0
  • Recombinant Human MGAT4A His-tag (rhMGAT4A) (Catalog # 11496-GT)
  • Cy5-Fuc labeled N2f (Cy5-N2f) (Catalog # GL304)
  • UDP-GlcNAc, 50 mM stock in 50% Ethanol
  • 17% SDS-PAGE Gel
  • Gel loading dye
  • Gel Imager with Cy5 fluorescent dye detection capability
  1. Dilute rhMGAT4A to 50 µg/mL with Assay Buffer.
  2. Create a Reaction Mix containing 0.02 µM Cy5-N2f and 1 mM UDP-GlcNAc in Assay Buffer.
  3. Combine 10 µL of rhMGAT4A and 10 µL of Reaction Mix. For a Control, combine 10 µL of Assay Buffer and 10 µL of Reaction Mix.
  4. Incubate reaction and control at 37 °C for 60 minutes.
  5. Add gel loading dye to each tube.
  6. Load half the volume of each reaction and control onto a 17% SDS-PAGE gel. Let samples migrate at least 80% down the gel before stopping.
  7. Acquire gel image and determine percent conversion of Cy5-N2f.
Per Reaction:
  • rhMGAT4A: 0.5 µg
  • Cy5-N2f: 0.2 pmol
  • UDP-GlcNAc: 0.5 mM

FAQs

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