Recombinant Human N-Acetylmannosamine Kinase/GNE Protein, CF

R&D Systems, part of Bio-Techne | Catalog # 8268-GK

R&D Systems, part of Bio-Techne
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8268-GK-050
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Key Product Details

Source

E. coli

Accession #

Q9Y223

Conjugate

Unconjugated

Applications

Enzyme Activity

View all N-Acetylmannosamine Kinase/GNE Proteins and Enzymes »

Product Specifications

Source

E. coli-derived human N-Acetylmannosamine Kinase/GNE protein
Thr406-Arg720, with an N-terminal Met and 6-His tag

Purity

>90%, by SDS-PAGE visualized with Silver Staining and quantitative densitometry by Coomassie® Blue Staining.

Endotoxin Level

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

N-terminal Sequence Analysis

Met & Leu410

Predicted Molecular Mass

34 kDa

SDS-PAGE

31-39 kDa, reducing conditions

Activity

Measured by its ability to phoshorylate N-Acetyl-D-Mannosamine.
The specific activity is >130 pmol/min/μg, as measured under the described conditions.

Formulation, Preparation and Storage

8268-GK
Formulation Supplied as a 0.2 μm filtered solution in Tris, NaCl and DTT.
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: N-Acetylmannosamine Kinase/GNE

Sialic acid modification of cell surface molecules is crucial for many biologic processes, such as cell adhesion (1), formation or masking of recognition determinants (2), and stabilization of glycoprotein structure of glycoproteins (3). Furthermore, sialic acids are over-expressed in many tumor cells (4) and are associated with a higher metastatic potential (5). The rate limiting enzyme of sialic acid biosynthesis is a bifunctional enzyme, UDP-N-acetylglucosamine-2-epimerase/N-acetylmannosamine kinase (GNE), that converts UDP-GlcNAc to ManNAc and subsequently phosphorylates the ManNAc to ManNAc 6-phosphate (6). The activity of GNE is regulated through feedback inhibition by the downstream product, CMP-sialic acid (7). In humans, GNE mutation causes two genetic diseases, sialuria (8) and hereditary inclusion body myopathy (HIBM). In sialuria, a mutation of either Arg-263 or Arg-266 in the epimerase domain impairs the feedback inhibition of GNE (9). HIBM is an autosomal recessive neuromuscular disorder caused by more than 60 different point mutations (10).The recombinant human N-Acetylmannosamine Kinase only contains the kinase domain (11). The kinase activity was measured using a phosphatase-coupled method (12).

References

  1. Varki, A. (2007) Nature 446:1023.
  2. Karlsson, K.-A. (1995) Curr. Opin. Struct. Biol. 5:622.
  3. Rens-Domiano, S. and Reisine, T. (1991) J. Biol. Chem. 266:20094.
  4. Roth, J. et al. (1988) Proc. Natl. Acad. Sci. U. S. A. 85:2999.
  5. Bresalier, R. S. et al. (1990) Cancer Res. 50:1299.
  6. Stasche, R. et al. (1997) J. Biol. Chem. 273: 24319.
  7. Kornfeld, S. et al. (1964) Proc. Natl.Acad. Sci. U.S.A. 52:371.
  8. Weiss, P. et al. (1989) J. Biol. Chem. 264:17635.
  9. Eisenberg, I. et al. (2001) Nat. Genet. 29:83.
  10. Huizing, M. and Krasnewich, D. M. (2009) Biochim. Biophys. Acta 1792:881.
  11. Martinez, J. et al. (2012) J. Biol. Chem. 287:13656.
  12. Wu, Z. (2011) PLoS ONE 6:e23172.

Long Name

Glucosamine (UDP-N-acetyl)-2-epimerase/N-acetylmannosamine Kinase

Alternate Names

DMRV, GLCNE, GNE, IBM2, NAcetylmannosamine Kinase, Uae1

Entrez Gene IDs

10020 (Human); 50798 (Mouse); 114711 (Rat)

Gene Symbol

GNE

UniProt

Q9Y223

Additional N-Acetylmannosamine Kinase/GNE Products

Product Documents for Recombinant Human N-Acetylmannosamine Kinase/GNE Protein, CF

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Product Specific Notices for Recombinant Human N-Acetylmannosamine Kinase/GNE Protein, CF

For research use only

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