GRF2 binding proteins and applications thereof
OwnerMichael F. Moran, Olga Ornatsky +1inventors
InventorsMichael F. Moran, Olga Ornatsky, Linda McBroom
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Abstract
Reagents and methods of use thereof regarding proteins and complexes of GRF2-interacting proteins (GRF2-IP) and proteins interaction with GRF2-IP, as well as polynucleotides encoding those proteins.
Claims22
An isolated protein complex including a combination of at least two proteins selected from the group consisting of GRF2, GRF2-Interacting Proteins, Ndr-Interacting Proteins, Skb1-Interacting Proteins, PP2C-Interacting Proteins, pICln-Interacting Proteins, 4.1SVWL2-Interacting Proteins, smD1-Interacting Proteins, and smD3-Interacting Proteins.
An isolated or recombinant protein having an amino acid sequence of a protein represented in Table 1, 2, 3, 4, 5, 6, 7, or 8 or a homolog thereof.
A method for identifying modulators of protein complexes, comprising the steps of:
- (i) forming a reaction mixture including a protein complex of at least two proteins selected from the group consisting of GRF2, GRF2-Interacting Proteins, Ndr-Interacting Proteins, Skb1-Interacting Proteins, PP2C-Interacting Proteins, pICln-Interacting Proteins, 4.1SVWL2-Interacting Proteins, smD1-Interacting Proteins, and smD3-Interacting Proteins,
- (ii) contacting the reaction mixture with a test agent, and
- (iii) determining the effect of the test agent for one or more activities selected from the group consisting of:
- (a) a change in the abundance of the protein complex;
- (b) a change in the activity of the complex;
- (c) a change in the activity of at least one member of the complex;
- (d) where the reaction mixture is a whole cell, a change in the intracellular localization of the complex or a component thereof;
- (e) where the reaction mixture is a whole cell, a change in the transcription level of a gene dependent on the complex;
- (f) where the reaction mixture is a whole cell, a change in the abundance of the product of a gene dependent on the complex;
- (g) where the reaction mixture is a whole cell, a change in the activity of the product of a gene dependent on the complex;
- and,
- (h) where the reaction mixture is a whole cell, a change in second messenger levels in the cell.
A method for identifying an agent which may modulate GRF2 dependent growth comprising:
- (i) forming a reaction mixture including a protein selected from the group consisting of GRF2-Interacting Proteins, Ndr-Interacting Proteins, Skb1-Interacting Proteins, PP2C-Interacting Proteins, pICln-Interacting Proteins, 4.1SVWL2-Interacting Proteins, smD1-Interacting Proteins, and smD3-Interacting proteins,
- (ii) contacting the reaction mixture with a test agent, and
- (iii) detecting the effect of the test agent for one or more activities selected from the group consisting of:
- (a) a change in the abundance of the protein complex;
- (b) a change in the activity of the complex;
- (c) a change in the activity of at least one member of the complex;
- (d) where the reaction mixture is a whole cell, a change in the intracellular localization of the complex or a component thereof;
- (e) where the reaction mixture is a whole cell, a change in the transcription level of a gene dependent on the complex;
- (f) where the reaction mixture is a whole cell, a change in the abundance of the product of a gene dependent on the complex;
- (g) where the reaction mixture is a whole cell, a change in the activity of the product of a gene dependent on the complex;
- and,
- (h) where the reaction mixture is a whole cell, a change in second messenger levels in the cell.
A method for inhibiting Ras-dependent proliferation of a cell comprising contacting the cell with an agent capable of inhibiting the activity of a member of the Ras signaling pathway.
A method for inhibiting Ras-dependent proliferation of a cell comprising contacting the cell with an inhibitor of a methyl transferase activity of Skb1.
A method for inhibiting Ras-dependent proliferation of a cell comprising contacting the cell with an inhibitor of a kinase activity of Skb 1.
15bis. A method for inhibiting Ras-dependent proliferation of a cell comprising contacting the cell with an agent that inhibits normal subcellular localization of Skb1.
A method for inhibiting Ras-dependent proliferation of a cell comprising contacting the cell with an inhibitor of a phosphatase activity of PP2C.
A method for inhibiting Ras-dependent proliferation of a cell comprising contacting the cell with an inhibitor of an activity of pICln.
A method for detecting aberrant GRF2-dependent signaling in a cell, comprising the step of screening the cell for one or more of:
- (i) altered levels of expression of a gene encoding a GRF2-Interacting Protein, an Ndr-Interacting Protein, an Skb1-Interacting Protein, a PP2C-Interacting Protein, a pICln-Interacting Protein, a 4.1SVWL2-Interacting Protein, an smD1-Interacting Protein, or an smD3-Interacting protein,
- (ii) altered levels of stability, post-translation modification, cellular localization and/or enzymatic activity of a GRF2-Interacting Protein, an Ndr-Interacting Protein, an Skb1-Interacting Protein, a PP2C-Interacting Protein, a pICln-Interacting Protein, a 4.1SVWL2-Interacting Protein, an smD1-Interacting Protein, or an smD3-Interacting protein, and
- (iii) altered levels of activity of a complex including a GRF2-Interacting Protein, an Ndr-Interacting Protein, an Skb1-Interacting Protein, a PP2C-Interacting Protein, a pICln-Interacting Protein, a 4.1SVWL2-Interacting Protein, an smD1-Interacting Protein, or an smD3-Interacting protein.
A method for inhibiting Ras-dependent proliferation of a cell comprising contacting the cell with an inhibitor of a kinase activity of Ndr.
A method for inhibiting Ras-dependent proliferation of a cell comprising contacting the cell with an agent that inhibits normal subcellular localization of Ndr.
Dependencies are inferred from the claim text.