• 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.  

• Activity

  Measured by the ability of the immobilized protein to support the adhesion of NIH?3T3 mouse embryonic fibroblast cells. When 5 x 10    ⁴ cells/well are added to recombinant human Nectin-4 coated plates (10 μg/mL with 100 μL/well), approximately >45% will adhere after 1 hour incubation at 37 °C.    
     Optimal dilutions should be determined by each laboratory for each application.  

• Source

  Mouse myeloma cell line, NS0-derived Gly27-Val351, with a C-terminal 6-His tag

• Accession #

• N-terminal Sequence    
  Analysis

  Gly27

• Structure / Form

  Monomer

• Predicted Molecular Mass

  35.6 kDa

• SDS-PAGE

  44 kDa, reducing conditions

Background: Nectin-4

Nectin-4 (gene name PVRL4, poliovirus receptor-like 4) is a 66 kDa type I transmembrane glycoprotein belonging to the Nectin family of Ig superfamily proteins (1). The Latin word necto means “to connect”, indicating the role of nectins in Ca²⁺?independent cell-cell adhesion (2). Nectin-4 forms homodimers in cis, followed by interactions in trans with Nectin-1 or -4 (1-3). Human Nectin-4 mRNA is normally expressed in the placenta, especially in endothelial cells, while in the mouse it is found in the embryo, lung, testis and brain (1, 4, 5). Human Nectin-4 cDNA encodes 510 amino acids (aa), including a 31 aa signal sequence, a 318 aa extracellular domain (ECD), a 21 aa transmembrane segment (TM), and a 140 aa cytoplasmic region. Nectin ECDs contain three Ig-like domains: an N?terminal V-type that mediates ligand binding, and two C2-type (1, 3). One Nectin?4 isoform lacks aa 412-436 in the cytoplasmic domain (1). In many human ductal breast or non-small cell lung carcinomas, Nectin?4 is upregulated and a soluble 43 kDa form is found in the plasma (4-6). This form is generated from the membrane protein via the action of TACE/ADAM-17 (6). The extracellular domain of human Nectin-4 shares 91%, 92%, 93%, 91% and 90% amino acid sequence homology with the corresponding regions of mouse, rat, canine, porcine and bovine Nectin-4, respectively. In forming adherens junctions, trans interactions of Nectin-4 initiate cell-cell interactions and recruit intracellular cadherins through afadin and other junctional proteins (1, 2). These interactions organize the actin cytoskeleton, strengthen attachment to basement membrane and promote further cell-cell connections (2, 7). In humans, mutation of Nectin-4 has been correlated with ectodermal dysplasia-syndactyly syndrome, indicating a role for Nectin-4 in human development (7).

• References:

1. Reymond, N. et al. (2001) J. Biol. Chem. 276:43205.

2. Takai, Y. et al. (2008) Nat. Rev. Mol. Cell Biol. 9:603.

3. Fabre, S. et al. (2002) J. Biol. Chem. 277:27006.

4. Fabre-Lafay, S. et al. (2007) BMC Cancer 7:73.

5. Takano, A. et al. (2009) Cancer Res. 69:6694.

6. Fabre-Lafay, S. et al. (2005) J. Biol. Chem. 280:19543.

7. Brancati, F. et al. (2010) Am. J. Hum. Genet. 87:265.

• Long Name:

  Poliovirus Receptor Related 4

• Entrez Gene IDs:

  81607 (Human); 71740 (Mouse)

• Alternate Names:

  EDSS1; Ig superfamily receptor LNIR; LNIR; LNIRnectin 4; Nectin4; Nectin-4; poliovirus receptor-related 4; poliovirus receptor-related protein 4; PRR4; PRR4DKFZp686K05193; PVRL4