c-Met, Recombinant, Canine (Hepatocyte Growth Factor Receptor, Scatter Factor Receptor, HGFR, SFR)

HGF R, also known as Met (from N-methyl-N'-nitro-N-nitrosoguanidine induced), is a glycosylated receptor tyrosine kinase that plays a central role in epithelial morphogenesis and cancer development. HGF R is synthesized as a single chain precursor which undergoes posttranslational proteolytic cleavage. This generates a mature HGF R that is a disulfide-linked dimer composed of a 50 kD extracellular a chain and a 145 kD transmembrane b chain. 1,2 The extracellular domain (ECD) contains a seven bladed b- propeller sema domain, a cysteine-rich PSI/MRS region, and four Ig-like E-set domains, while the cytoplasmic region includes a tyrosine kinase domain. 3 The sema domain, which is formed by both the a and b chains of HGF R, mediates both ligand binding and receptor dimerization. 3,4 Ligand-induced tyrosine phosphorylation in the cytoplasmic region activates the kinase domain and provides docking sites for multiple SH2-containing molecules. 5,6 HGF stimulation induces HGF R downregulation via internalization and proteasome-dependent degradation. 7 In the absence of ligand, HGF R forms noncovalent complexes with a variety of membrane proteins including CD44v6, CD151, EGF R, Fas, integrin a 6/b4, plexins B1, B2, and B3, and MSP R/Ron. 8-15 Ligation of one complex component triggers activation of the other, followed by cooperative signaling effects. 8-15 Formation of some of these heteromeric complexes is a requirement for epithelial cell morphogenesis and tumor cell invasion. 8,12,13 HGF released from neighboring mesenchymal cells stimulates HGF R on undifferentiated epithelium and induces epithelial cell scattering and branching tubulogenesis. 16 Genetic polymorphisms, chromosomal translocation, overexpression, and additional splicing and proteolytic cleavage of HGF R have been described in a wide range of cancers. 1 Within the ECD, canine HGF R shares 85%-88% aa sequence identity with human, mouse and rat HGF R.\n\nSource: \nHuman CD33 signal peptide (Met 1-Ala 16), Canine HGF R, (Glu 25-Leu 935), HHHHHH; A DNA sequence encoding the signal peptide from human CD33 joined with the extracellular domain of canine HGF R (Glu 25-Leu 935; Accession # Q75ZY9) was fused with a 6X histidine tag at the C-terminus. The chimeric protein was expressed in a mouse myeloma cell line, NS0.\n\nMolecular Mass: \nThe mature recombinant canine HGF R/His is a disulfide-linked heterodimer. Based on N-terminal amino acid sequencing, the recombinant canine HGF R/His has two N-termini starting at Glu 25 and Ser 309 that correspond to the a and b chains, respectively. The predicted molecular mass for the a chain is approximately 32.6 kD, and that for the b chain is approximately 70.1 kD. As a result of glycosylation, in SDS-PAGE, the recombinant protein migrates as an approximately 130-150 kD protein under non-reducing conditions, and as approximately 42-47 kD (a-chain) and 90-100 kD (b?chain) proteins under reducing conditions.\n\nEndotoxin Level: \n< 1.0 EU per 1 microg of the protein as determined by the LAL method.\n\nActivity: \nMeasured by its ability to bind rcaHGF with an estimated Kd < 0.8 nm\n\nReoconstitution: \nIt is recommended that sterile PBS be added to the vial to prepare a working stock solution of no less than 100ug/ml. The carrier-free protein should be used immediately upon reconstitution to avoid losses in activity due to non-specific binding to the inside surface of the vial. For long term storage as a dilute solution, a carrier protein (e.g. 0.1% HSA or BSA) should be added to the vial.\n\nStorage and Stability:\nLyophilized samples are stable for up to twelve months from date of receipt at -20 degrees C. Upon reconstitution, this protein, in the presence of a carrier protein, can be stored under sterile conditions at 2-8 degrees C for one month or at -20 degrees C in a manual defrost freezer for three months without detectable loss of activity. Avoid repeated freeze-thaw cycles.

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SPECIFICATIONS

Catalog Number

M3007-13C

Size

50ug

Form

Supplied as a lyophilized powder from a 0.2um filtered solution in PBS.

Purity

(same/more than) 95%, as determined by SDS-PAGE and visualized by silver stain.

References

1. Birchmeier, C. et al., 2003, Nat. Rev. Mol. Cell Biol. 4:915. \n2. Corso, S. et al., 2005, Trends Mol. Med. 11:284. \n3. Gherardi, E. et al., 2003, Proc. Natl. Acad. Sci. USA 100:12039. \n4. Kong-Beltran, M. et al., 2004, Cancer Cell 6:75. \n5. Naldini, L. et al., 1991, Mol. Cell. Biol. 11:1793. \n6. Ponzetto, C. et al., 1994, Cell 77:261. \n7. Jeffers, M. et al., 1997, Mol. Cell. Biol. 17:799. \n8. Orian-Rousseau, V. et al., 2002, Genes Dev. 16:3074. \n9. Klosek, S.K. et al., 2005, Biochem. Biophys. Res. Commun. 336:408. \n10. Jo, M. et al., 2000, J. Biol. Chem. 275:8806. \n11. Wang, X. et al., 2002, Mol. Cell 9:411. \n12. Trusolino, L. et al., 2001, Cell 107:643. \n13. Giordano, S. et al., 2002, Nat. Cell Biol. 4:720. \n14. Conrotto, P. et al., 2004, Oncogene 23:5131. \n15. Follenzi, A. et al., 2000, Oncogene 19:3041. \n16. Sonnenberg, E. et al., 1993, J. Cell Biol. 123:223.