Recombinant Rat GFR alpha-1 Fc Chimera Protein Summary
Details of Functionality
Measured by its ability to confer GDNF responsiveness to PC‑12 rat adrenal pheochromocytoma cells using a neurite outgrowth assay. Treanor, J. et al. (1996) Nature 392:80. The ED50 for this effect is approximately 0.3 µg/mL in the presence of 25 ng/mL of Recombinant Rat GDNF (Catalog # 512-GF). Measured by its binding ability in a functional ELISA. Immobilized Recombinant Rat GDNF (Catalog # 512-GF) at 1 μg/mL binds Recombinant Rat GFR alpha ‑1/GDNF R alpha ‑1 Fc Chimera with an apparent KD <5 nM.
Mouse myeloma cell line, NS0-derived rat GFR alpha-1/GDNF R alpha-1 protein
Rat GFR alpha -1 (Asp25-Leu445) Accession # Q62997
>90%, by SDS-PAGE under reducing conditions and visualized by silver stain
<0.10 EU per 1 μg of the protein by the LAL method.
74 kDa (monomer). Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Glial cell line-derived growth factor (GDNF), neurturin (NTN) and persephin, distant members of the TGF‑ beta superfamily, are neurotrophic factors for a variety of neuronal populations in the central and peripheral nervous systems. The bioactivities of GDNF and NTN are mediated through a receptor complex composed of the non ligand-binding signaling subunit (c-Ret receptor tyrosine kinase) and either of two ligand binding subunits (GDNF receptor alpha - (GFR alpha -1) or GFR alpha -2). GFR alpha -1 and -2 are members of a family of at least four cysteine‑rich glycosyl-phosphatidylinositol (GPI)‑linked cell surface proteins that share conserved placements of many of their cysteine residues. Binding of GDNF to membrane-associated GFR alpha -1 or GFR alpha -2 initiates the association with and activation of the Ret tyrosine kinase. Soluble GFR alpha s released enzymatically from the cell surface-associated protein with phosphatidylinositol phospholipase C, as well as recombinantly produced soluble GFR alpha -1, can also bind with high-affinity to GDNF and trigger the activation of Ret tyrosine kinase.
Rat GFR alpha -1 cDNA encodes a 468 amino acid (aa) residue protein with an N-terminal 24 aa residue hydrophobic signal peptide. Like other GPI-linked proteins, rat GFR alpha -1 has a C-terminal hydrophobic region which is preceded by a three aa residue (ASS) GPI-binding site. Human GFR alpha -1 shares 93% amino acid identity with rat GFR alpha -1. The expression of the various GFR alpha s are differentially regulated in the central and peripheral nervous system, suggesting complementary roles for the GFR alpha s in mediating the activities of the GDNF family of neurotrophic factors.
Thompson, J. et al. (1998) Mol. Cell Neurosci. 11:117.
Trupp, M. et al. (1998) Mol. Cell Neurosci. 11:47.
Baloh, R.H. et al. (1998) Proc. Natl. Acad. Sci. USA 95:5801.
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