Glypican 1, Recombinant, Human (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, with one subfamily containing Glypicans 1, 2, 4 and 6, and another subfamily containing Glypicans 3 and 5 (3, 5). Human Glypican 1 (GPC-1) is synthesized as a 558 amino acid (aa) preproprecursor that contains a 23 aa signal sequence, a 507 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 are potentially two heparan sulfate (HS) modifications on GPC-1 that could contribute to a native molecular weight of approximately 200kD (7, 8, 9). Mature human GPC-1 shares 91% aa identity with mature mouse GPC-1. There are two potential splice variants of human GPC-1. Both show an alternate start site at Met73, while one has an additional 65 aa substitution for the C-terminal 264 amino acids (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). This is accomplished 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 contract, 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\nSource: A DNA sequence encoding the mature human Glypican 1 (Asp 24-Ser 530; Accession # P35052) was fused at the C-terminus to a six-histidine tag. The protein was expressed in a mouse myeloma cell line, NS0.\n\nDefinition: Measured by its ability to bind FGF-basic in a functional ELISA.\n\nApplications: \nSuitable for use in Western Blot and ELISA. Other applications not tested.\n\nRecommended Dilution:\nWestern Blot: 56.8kD protein migrates as an ~ 60kD protein under reducing conditions.\nELISA: 1 at 3ug/ml (100ul/well) will bind rhFGF-basic with an apparent KD (same/less than)1 nM.\nOptimal dilutions to be determined by the researcher.\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.

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SPECIFICATIONS

Catalog Number

G8235-46

Size

50ug

Applications

ELISA, WB

Reactivities

Hum

Form

Supplied as a lyophilized powder in PBS. Reconstitute with sterile PBS to (same/more than) 0.1mg/ml.

Purity

(same/more than) 95%, as determined by SDS-PAGE under reducing conditions and visualized by silver stain.\nEndotoxin (same/less than) 1EU.

References

1. Song, H.H. & J. Filmus (2002) Biochim. Biophys. Acta 1573:241. 11. GenBank Accession # EAW71183.\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 # P35052. 16. Selleck, S.B. (2006) SciSTKE, April 4:pe17\n7. David, G. et al. (1990) J. Cell Biol. 111:3165. 17. Qiao, D. et al. (2003) J. Biol. Chem. 278:16045.\n8. Lories, V. et al. (1992) J. Biol. Chem. 267:1116. 18. Su, G. et al. (2006) Am. J. Pathol. 168:2014.\n9. Lories, V. et al. (1989) J. Biol. Chem. 264:7009. 19. Allen, B.L. & A.C. Rapraeger (2003) J. Cell Biol. 163:637.\n10. GenBank Accession # EAW71184.