Glypican 1, Recombinant, Mouse (Glypiated Proteoglycan 1)

The Glypicans (glypiated proteoglycans) are a small multigene family of GPI-linked proteoglycans that play a key role in growth factor signaling (1, 2, 3, 4). There are currently six known mammalian Glypicans. They all share a common-sized protein core of 60-70kD, an N-terminus which likely forms a compact globular domain, 14 conserved cysteines that form multiple intrachain disulfide bonds, and a number of C-terminal N- and O-linked carbohydrate attachment sites. Based on exon organization and the location of O-linked glycosylation sites, at least two subfamilies of Glypicans are known. One subfamily contains Glypicans 1, 2, 4 and 6, while another subfamily contains Glypicans 3 and 5 (3, 5). Mouse Glypican 1 (GPC-1) is synthesized as a 557 amino acid (aa) preproprecursor that contains a 23 aa signal sequence, a 506 aa mature segment, and a 28 aa C-terminal prosegment (6, 7). There are two potential N-linked, and four potential O-linked sites for glycosylation or glycanation. There is at least one heparan sulfate (HS) modification on GPC-1 that contributes to a native molecular weight of approximately 110kD (8, 9). Mature mouse GPC-1 shares 91% and 98% aa identity with mature human and rat GPC-1, respectively. There are two potential splice variants of mouse GPC-1. The first is truncated and shows a seven amino acid substitution for the first 294 amino acids; the second reveals an alternate start site at Met73 (10, 11). Cells known to express GPC-1 include neurons, smooth and skeletal muscle cells, keratinocytes, osteoblasts, Schwann cells, immature dendritic cells, and tumor, plus tumor-associated vascular endothelial cells (8, 9, 12-15). The function of GPC-1 is complex and varied. As a proteoglycan, it appears to make use of its HS adduct to impact select growth factor activity (16). GPC-1 accomplishes its co-receptor role by having juxtramembrane HS attachment sites and a flexible, GPI-linkage (17). Recent data suggests GPC-1 and sulfation enzymes may collaborate to regulate FGF signaling. HS modules that are rich in 2-O- and 6-O- sulfate upregulate FGF-2 activation of FGFR1c (18). Similarly, FGF-1 requires both 2-O- and 6-O-sulfation to bind to FGFR2c and 3c. By contrast, FGF-1 requires no sulfation to bind to FGFR2b, and FGF-8b needs only 6-O-sulfation to activate FGFR3c. Thus, many FGF receptor isoform specific effects may be attributed to an interaction between Glypican family members and the cell sulfation system (19).\n\nA DNA sequence encoding the mature mouse Glypican 1 (Asp 24-Ser 529; Accession # Q9QZF2) was fused to a six-hisidine tag at the C-terminus. The protein was expressed in a mouse myeloma cell line, NS0.\n\nCalculated molecular mass of 56.4kD. As a result of glycosylation, the recombinant protein migrates as an ~ 57-62kD protein in SDS-PAGE under reducing conditions.\n\nActivity: Measured by its ability to bind FGF-basic in a functional ELISA.\n\nStorage and Stability:\nLyophilized powder may be stored at 4 degrees C for short-term only. Reconstitute to nominal volume by adding sterile PBS and store at -20 degrees C. Reconstituted product is stable for 12 months at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

Prices direct from United States Biological

Quick response times

Exclusive Biosave savings/discounts

SPECIFICATIONS

Catalog Number

G8235-40M

Size

50ug

Form

Supplied as a lyophilized powder in PBS. Reconstitute with PBS to (same/more than)100ug/ml.

Purity

(same/more than) 90%, as determined by SDS-PAGE under reducing conditions and visualized by silver stain.\nEndotoxin: (same/less than) 1EU/1ug.

References

. Song, H.H. & J. Filmus (2002) Biochim. Biophys. Acta 1573:241. 11. GenBank Accession # EDL39993.\n2. Fransson, L-A. et al. (2004) Cell. Mol. Life Sci. 61:1016. 12. Chernousov, M.A. et al. (2006) J. Neurosci. 26:508.\n3. De Cat, B. & G. David (2001) Semin. Cell Dev. Biol. 12:117. 13. Wegrowski, Y. et al. (2006) Clin. Exp. Immunol. 144:485.\n4. Lamoureux, F. et al. (2007) BioEssays 29:758. 14. Qiao, D. et al. (2003) J. Biol. Chem. 278:16045.\n5. Veugelers, M. et al. (1999) J. Biol. Chem. 274:26968. 15. Kayed, H. et al. (2006) Int. J. Oncol. 29:1139.\n6. GenBank Accession # Q9QZF2. 16. Selleck, S.B. (2006) SciSTKE April 4:pe17\n7. Watanabe, K. et al. (1995) J. Cell Biol. 130:1207. 17. Qiao, D. et al. (2003) J. Biol. Chem. 278:16045.\n8. Litwick, E.D. et al. (1994) J. Neurosci. 14:3713. 18. Su, G. et al. (2006) Am. J. Pathol. 168:2014.\n9. Litwick, E.D. et al. (1998) Dev. Dyn. 211:72. 19. Allen, B.L. & A.C. Rapraeger (2003) J. Cell Biol. 163:637.\n10. GenBank Accession # EDL39991.