• Purity

  >95%, by SDS-PAGE under reducing conditions and visualized by silver stain

• Endotoxin Level

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

• Activity

  Measured by its ability to inhibit Recombinant Human Midkine (Catalog # ) induced neurite outgrowth in E16-E18 rat embryonic cortical neurons. When 5 x 10    ⁴ neurons per well are added to a plate pre-incubated with serial dilutions of rhNeuroglycan-C and 1 μg/mL of rhMidkine, the neurite outgrowth promotion is significantly inhibited in a dose dependent manner with an effect at    
  0.6-1.2 μg/mL.

• Source

  Chinese Hamster Ovary cell line, CHO-derived Val31-Gln420, with a C-terminal 6-His tag

• Accession #

• N-terminal Sequence    
  Analysis

  Val31

• Predicted Molecular Mass

  41.9 kDa

• SDS-PAGE

  80-100 kDa, reducing conditions

Background: Neuroglycan C/CSPG5

Neuroglycan C (NGC; also CSPG5 and CALEB) is a 120 - 150 kDa type I transmembrane glycoprotein and member of the neuregulin family of proteins (1 ‑ 2). Depending on its expression profile, NGC may be a glycoprotein of 120 kDa, or a chondroitin sulfate (CS) proteoglycan of 150 kDa (2 - 3). Human NGC is synthesized as a 566 amino acid (aa) precursor that contains a 30 aa signal sequence, a 393 aa extracellular domain (ECD), a 21 aa transmembrane segment, and a 122 aa cytoplasmic region. The ECD contains one CS attachment domain (aa 34 - 272), with CS attachment at Ser117, one EGF-like domain (aa 371 - 413), two potential sites for N-linked glycosylation, and twelve potential sites for O-linked glycosylation (4). Splicing variants produce three isoforms for human NGC. Isoform 1 is the long form. Isoform 2 has a deletion of aa 487 ‑ 513, while isoform 3 has an alternative start site at Met139 and the same deletion. Phosphorylation likely occurs at Ser249, and proteolysis generates a 75 kDa soluble fragment (5). Over aa 31 - 420, human NGC shares 84% aa identity with mouse NGC. NGC is expressed in nervous tissue and is found on retinal ganglion cells, cerebellar Purkinje cells and hippocampal neurons (6). NGC may function as a growth and differentiation factor involved in neuritogenesis. One study shows that the recombinant ectodomain of NGC core protein enhances neurite outgrowth from rat neocortical neurons in culture via phosphatidylinositol 3-kinase and protein kinase C signaling pathways (7). Another study states that NGC is a novel component of midkine receptors, a heparin-binding growth factor that promotes cell attachment and process extension in oligodendroglial precursor-like cells (3). NGC also acts as a growth factor by directly binding ERbB3 tyrosine kinase and transactivating ErbB2 (1).

• References:

  
     

1. Kinugasa, Y. et al. (2004) Biochem. Biophys. Res. Commun 321:1045.

2. Yasuda, Y. et al. (1998) Neurosci. Res. 32:313.

3. Ichihara-Tanaka, K. et al. (2006) J. Biol. Chem. 281:30857.

4. Aono, S. et al. (2004) J. Biol. Chem. 279:46536.

5. Shuo, T. et al. (2007) J. Neurochem. 102:1561.

6. Aono, S. et al. (2006) J. Neurosci. Res.83:110.

7. Nakanishi, K. et al. (2006) J. Biol. Chem. 281:24970.

• Entrez Gene IDs:

  10675 (Human); 29873 (Mouse); 50568 (Rat)

• Alternate Names:

  CALEB; chondroitin sulfate proteoglycan 5 (neuroglycan C); chondroitin sulfate proteoglycan 5; CSPG5; MGC44034; Neuroglycan C; NGC; NGCAcidic leucine-rich EGF-like domain-containing brain protein