• Purity

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

• Endotoxin Level

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

• Activity

  Measured by the ability of the immobilized protein to support the adhesion of SVEC4‑10 mouse vascular endothelial cells. When 4 x 10    ⁴ cells/well are added to rhNidogen-1 coated plates (30 µg/mL with 100 µL/well), approximately 40-75% will adhere after one hour at 37 °C.    
     Optimal dilutions should be determined by each laboratory for each application.  

• Source

  Mouse myeloma cell line, NS0-derived Leu29-Lys1114 (Gln1113Arg), with an N-terminal 9-His tag

• Accession #

• N-terminal Sequence    
  Analysis

  His

• Predicted Molecular Mass

  120 kDa

• SDS-PAGE

  130 kDa, reducing conditions

Background: Nidogen-1/Entactin

Nidogen-1 (also entactin) is a 150 kDa, secreted, monomeric glycoprotein that serves as a major linking component of basement membranes (1 - 4). It is synthesized as a 1247 amino acid (aa) precursor with a 28 aa signal sequence and a 1219 aa mature protein. The molecule is modular in structure with five distinct regions. There are three globular domains (G1-3) separated by a mucin region and an extended rod-shaped segment (5 - 7). The N-terminal globular domain (G1) is 200 aa in length and seemingly unrelated to any known motif (8). The mucin region is nearly 160 aa in length and presumably O-glycosylated (2, 8). G2 and G3 are both approximately 300 aa in length. G2 is described as a Nidogen ( beta -barrel) domain, while C-terminal G3 assumes a beta -propeller configuration (1). The 250 aa rod-shaped segment has multiple EGF-like motifs and two thyroglobulin type 1 domains. Functionally, G1 is reported to bind type IV collagen (2, 7). The mucin region contains a short peptide that ligates alpha ₃ beta ₁ integrins (9, 10). G2 interacts with perlecan, and an RGD motif in the rod-shaped segment serves as a binding site for alpha _v beta ₃ integrins (9, 10). Finally, G3 is associated with laminin binding (2, 7). As a full-length molecule, the multiple extracellular matrix-binding sites of Nidogen-1 are well positioned to serve as anchor sites for basement membrane molecules. Nidogen-1 also undergoes proteolytic processing by at least two MMPs, MMP7 and MMP19 (10, 11). While this destroys the integrity of Nidogen-associated matrices, it also generates peptide fragments that are capable of inducing neutrophil chemotaxis and phagocytosis (10). Nidogen-2 is related to Nidogen-1 (≈ 50% aa identity) and shares many of the same adhesive properties as Nidogen-1 (12). Both bind perlecan plus collagens I and IV. Nidogen-2, however, does not bind fibulin-1 or 2, and shows only modest interaction with laminin. Thus, although coexpressed, Nidogen-2 serves as only a partial substitute for Nidogen-1 (2, 12). Human Nidogen-1 is 85% aa identical to both mouse and rat Nidogen-1, and 88% aa identical to canine Nidogen-1.

• References:

1. Hohenester, E. and J. Engel (2002) Matrix Biol. 21:115. 

2. Miosge, N. et al. (2001) Histochem. J. 33:523.

3. Charonis, A. et al. (2005) Curr. Med. Chem. 12:1495.

4. Timpl, R. and J.C. Brown (1996) BioEssays 18:123. 

5. Nagayoshi, T. et al. (1989) DNA 8:581. 

6. Zimmerman, K. et al. (1995) Genomics 27:245. 

7. Fox, J.W. et al. (1991) EMBO J. 10:3137.

8. Mayer, U. et al. (1995) Eur. J. Biochem. 227:681.

9. Gresham, H.D. et al. (1996) J. Biol. Chem. 271:30587.

10. Dong, L-J. et al. (1995) J. Biol. Chem. 270:15383.

11. Titz, B. et al. (2004) Cell. Mol. Life Sci. 61:1826.

12. Kohfeldt, K. et al. (1998) J. Mol. Biol. 282:99.

• Entrez Gene IDs:

  4811 (Human)

• Alternate Names:

  enactin; Entactin; Entactin-1; NID1; NID-1; NIDentactin; nidogen (enactin); nidogen 1; Nidogen1; Nidogen-1