Isolated FGF receptor
Disclosed herein are compositions and methods which are useful in the identification and isolation of components involved in transmembrane receptor-mediated signaling. Such components include the receptors themselves (e.g., tyrosine kinase receptors, cytokine receptors and tyrosine phosphatase receptors), as well as ligands which bind the receptors and modulators of the downstream intracellular catalytic event which characterizes receptor-mediated signalling. Two novel ligands for the FGF receptor and the nucleotide sequences encoding them are also described.
1. A method for identifying an FGF receptor ligand comprising:
a) providing a cDNA expression library from an organism of interest;
b) providing a population of cells that express a DNA sequence encoding a heterologous FGF receptor that comprises an extracellular domain, a transmembrane domain, and an intracellular domain that is characterized by protein tyrosine kinase activity;
c) transforming the population of cells with the cDNA expression library; and
d) detecting protein kinase activity in clonally-derived cells, wherein elevated FGF receptor tyrosine kinase activity indicates the presence of an FGF receptor ligand.
2. The method according to claim 1 , wherein the cell is a yeast cell.
3. The method according to claim 1 , wherein the DNA sequence encoding a heterologous FGF receptor is carried on a CEN-based plasmid.
4. The method according to claim 1 , wherein the DNA sequence encoding a heterologous FGF receptor is inserted into a chromosome.
5. A method for identifying an FGF receptor ligand comprising:
a) providing a cDNA expression library from an organism of interest under the control of an inducible promoter;
b) providing a population of cells that express a DNA sequence encoding a heterologous FGF receptor that comprises an extracellular domain, a transmembrane domain, and an intracellular domain that is characterized by protein tyrosine kinase activity;
c) transforming the population of cells with the cDNA expression library; and
d) detecting protein kinase activity in clonally-derived cells, wherein elevated FGF receptor tyrosine kinase activity indicates the presence of an FGF receptor ligand.
6. The method according to claim 5 , wherein the cell is a yeast cell.
7. The method according to claim 5 , wherein the DNA sequence encoding a heterologous FGF receptor is carried on a CEN-based plasmid.
8. The method according to claim 5 , wherein the DNA sequence encoding a heterologous FGF receptor is inserted into a chromosome.
9. The method according to claim 5 , wherein the DNA sequence encoding a heterologous FGF receptor is constitutively expressed.
10. A method for identifying an FGF receptor ligand comprising:
a) providing a cDNA expression library from an organism of interest;
b) providing a population of cells that constitutively express a DNA sequence encoding a heterologous FGF receptor that comprises an extracellular domain, a transmembrane domain, and an intracellular domain that is characterized by protein tyrosine kinase activity;
c) transforming the population of cells with the cDNA expression library; and
d) detecting protein kinase activity in clonally-derived cells, wherein elevated FGF receptor tyrosine kinase activity indicates the presence of an FGF receptor ligand.
11. The method according to claim 10 , wherein the cell is a yeast cell.
12. The method according to claim 10 , wherein the DNA sequence encoding a heterologous FGF receptor is carried on a CEN-based plasmid.
13. The method according to claim 10 , wherein the DNA sequence encoding a heterologous FGF receptor is inserted into a chromosome.
14. The method according to claim 10 , wherein the cDNA expression library having polynucleotide inserts under the control of an inducible promoter.