• Purity

  >80%, 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 neurite outgrowth of dissociated E13 chick embryonic dorsal root ganglia (DRG) neurons. Able to significantly inhibit neurite outgrowth when immobilized as a 3 μL droplet containing 200 ng on a nitrocellulose-coated microplate.

• Source

  Chinese Hamster Ovary cell line, CHO-derived

  Human Nogo-A (Val566 - Phe748) & (Thr569 - Phe748) Accession # Q9NQC3	IEGRMD	Human IgG1 (Pro100 - Lys330)
  N-terminus		C-terminus

• Accession #

• N-terminal Sequence    
  Analysis

  Val566 & Thr569

• Structure / Form

  Disulfide-linked homodimer

• Predicted Molecular Mass

  46 kDa (monomer)

• SDS-PAGE

  70-90 kDa, reducing conditions

Background: Nogo-A

Human Nogo-A (also reticulon-4) is a member of the reticulon family of transmembrane proteins. This family is characterized by the presence of a nonsignal sequence-containing N-terminus, a topologically conserved 200 amino acid (aa) C-terminus that contains two transmembrane domains with an ER-retention motif, and a punctate intracellular distribution within the ER that is reminescent of a reticulum (1 - 4). In human, Nogo exists in five isoforms (5 - 7). The full length human form (Nogo-A) is 1192 aa in length and contains a 1018 aa N-terminus, a 21 aa transmembrane segment, a 94 aa connecting “l(fā)oop”, a second 21 aa transmembrane segment, and a 38 aa C-terminus. Three areas are of particular interest. One is a stretch of 66 aa within the 94 aa connecting loop. This segment is reported to bind to the GPI-linked Nogo receptor/p75 complex on axons and induce growth cone collapse (8 - 10). Two other areas in the N-terminus have also been discovered to have bioactivity (8, 11, 12). Based on rat, aa 57 - 184 in human (aa 59 - 172 in rat) should block fibroblast spreading, while aa 566 - 748 in human (aa 544 - 725 in rat) block neurite outgrowth and block fibroblast spreading (8, 12, 13). The exact topology of Nogo-A is unclear. The N- and C-termini may be extracellular with the “l(fā)oop” region intracellular, or the situation could be reversed (13 - 15). Alternatively, the loop region and N-terminus may be on the same side of the membrane (3, 8). The four additional isoforms are shorter than Nogo-A (199 aa [Nogo-C], 373 aa [Nogo-B], 392 aa and 986 aa, respectively) (7). Although highly divergent, all contain the same C-terminal stretch, aa 1005 - 1192. Both Nogo-B and C are reported to complex with Nogo-A (16). Notably, Nogo-A is expressed in neurons, endothelial cells. oligodendrocytes, fibroblasts and myoblasts (12, 16 - 18). Human Nogo-A is 78% aa identical to mouse and rat Nogo-A overall, with 98% aa identity in the loop region and approximately 80% aa identity in the aa 566 - 748 segment.

• References:

1. Oertle, T. et al. (2003) FASEB J. 17:1238.

2. GrandPre, T. et al. (2000) Nature 403:439.

3. Yan, R. et al. (2006) Cell. Mol. Life Sci. 63:877.

4. Teng, F.Y.H. et al. (2004) J. Neurochem. 89:801.

5. Chen, M.S. et al. (2000) Nature 403:434.

6. Morris, N.J. et al. (1999) Biochim. Biophys. Acta 1450:68.

7. GenBank Accession # Q9NQC3.

8. Oertle, T. et al. (2003) J. Neurosci. 23:5393.

9. Fournier, A.E. et al. (2001) Nature 409:341.

10. Wang, K.C. et al. (2002) Nature 420:74.

11. Prinjha, R. et al. (2000) Nature 403:384.

12. Dodd, D.A. et al. (2005) J. Biol. Chem. 280:12494.

13. Li, M. et al. (2004) Eur. J. Biochem. 271:3512.

14. Huber, A.B. and M.E. Schwab (2000) Biol. Chem. 381:407.

15. Ng, C.E.L. and B.L. Tang (2002) J. Neurosci. Res. 67:559.

16. Dodd, D.A. et al. (2005) J. Biol. Chem. 280:12494.

17. Wang, X. et al. (2002) J. Neurosci. 22:5505.

18. Acevedo, L. et al. (2004) Nat. Med. 10:382.

• Long Name:

  Reticulon 4A

• Entrez Gene IDs:

  57142 (Human); 68585 (Mouse); 83765 (Rat)

• Alternate Names:

  NI220; NogoA; Nogo-A; RTN4; RTN4A