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You are here:Home » Molecular Biology » MB-Binding Proteins » Glypican 1, Recombinant, Human (Glypiated Proteoglycan 1)

Glypican 1, Recombinant, Human (Glypiated Proteoglycan 1)

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Specifications

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).
Catalog #G8235-46
Source 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.
Definition Measured by its ability to bind FGF-basic in a functional ELISA.
ApplicationsSuitable for use in Western Blot and ELISA. Other applications not tested.
Recommended DilutionWestern Blot: 56.8kD protein migrates as an ~ 60kD protein under reducing conditions.
ELISA: 1 at 3ug/ml (100ul/well) will bind rhFGF-basic with an apparent KD 1 nM.
Optimal dilutions to be determined by the researcher.
Storage and StabilityLyophilized powder may be stored at 4°C for short-term only. Reconstitute to nominal volume by adding sterile PBS and store at -20°C. Reconstituted product is stable for 12 months at -20°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.
Molecular Weight56.8kD
SourceRecombinant Human
Purity95%, as determined by SDS-PAGE under reducing conditions and visualized by silver stain.
Endotoxin 1EU.
ConcentrationAs reported
FormSupplied as a lyophilized powder in PBS. Reconstitute with sterile PBS to 0.1mg/ml.
Important NoteThis product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.


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