COX 1-interacting proteins and use thereof
AssigneeMyriad Genetics, Inc.
InventorDaniel Albert Wettstein
Life of this patent
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Abstract
Protein complexes are provided comprising COX1 and one or more proteins selected from the group consisting of THR S14 and Opa1. The protein complexes are useful in screening assays for identifying compounds effective in modulating the protein complexes and in treating and/or preventing diseases and disorders associated with COX1 and its interacting partner proteins. In addition, methods of detecting the protein complexes and modulating the functions and activities of the protein complexes or interacting members thereof are also provided.
Claims38
An isolated protein complex having a first protein which is COX1 or a homologue or derivative or fragment thereof interacting with a second protein which is a protein selected from the group consisting of THR S14 and Opa1 or a homologue or derivative or fragment thereof.
An isolated protein complex comprising a first protein interacting with a second protein, wherein:
- (a) said first protein is selected from the group consisting of
- (i) COX1,
- (ii) a COX1 fragment capable of interacting with a protein selected from the group consisting of THR S14 and Opa1, and
- (iii) a fusion protein containing COX1 or said COX1 fragment;
- and
- (b) said second protein is selected from the group consisting of
- (1) a protein selected from the group consisting of THR S14 and Opa1,
- (2) a fragment of a protein selected from the group consisting of THR S14 and Opa1 and capable of interacting with COX1, and
- (3) a fusion protein containing a protein selected from the group consisting of THR S14 and Opa1 or said fragment.
A fusion protein having a first polypeptide covalently linked to a second polypeptide, wherein said first polypeptide is COX1 or a homologue or fragment thereof, and wherein said second polypeptide is a protein selected from the group consisting of THR S14 and Opa1 or a homologue or fragment thereof.
A method for selecting modulators of an interaction between a first protein and a second protein,
- (a) said first protein being selected from the group consisting of
- (i) COX1,
- (ii) a COX1 homologue having an amino acid sequence at least 90% identical to that of COX1 and capable of interacting with a protein selected from the group consisting of THR S14 and Opa1,
- (iii) a COX1 fragment capable of interacting with a protein selected from the group consisting of THR S14 and Opa1, and
- (iv) a fusion protein containing COX1, said COX1 homologue or said COX1 fragment;
- and
- (b) said second protein being selected from the group consisting of
- (1) THR S14 and Opa1,
- (2) a homologue of a protein selected from the group consisting of THR S14 and Opa1 having an amino acid sequence at least 90% identical to that of said protein and capable of interacting with COX1,
- (3) a fragment of a protein selected from the group consisting of THR S14 and Opa1 and capable of interacting with COX1, and
- (4) a fusion protein containing a protein selected from the group consisting of THR S14 and Opa1, said protein homologue or said protein fragment, said method comprising:
- contacting said first protein with said second protein in the presence of a test compound;
- and
- detecting the interaction between said first protein and said second protein.
A method for selecting modulators of an interaction between a first polypeptide and a second polypeptide, said first polypeptide being COX1 or a homologue or fragment thereof and said second polypeptide being a protein selected from the group consisting of THR S14 and Opa1 or a homologue or fragment thereof, said method comprising:
- providing in a host cell a first fusion protein having said first polypeptide, and a second fusion protein having said second polypeptide, wherein a DNA binding domain is fused to one of said first and second polypeptides while a transcription-activating domain is fused to the other of said first and second polypeptides;
- providing in said host cell a reporter gene, wherein the transcription of the reporter gene is controlled by the interaction between the first polypeptide and the second polypeptide;
- allowing said first and second fusion proteins to interact with each other within said host cell in the presence of a test compound;
- and
- determining the expression of said reporter gene.
A method for selecting compounds capable of interfering with the interaction between a first protein and a second protein, wherein
- (a) said first protein is selected from the group consisting of
- (i) COX1,
- (ii) a COX1 homologue having an amino acid sequence at least 90% identical to that of COX1 and capable of interacting with a protein selected from the group consisting of THR S14 and Opa1,
- (iii) a COX1 fragment capable of interacting with a protein selected from the group consisting of THR S14 and Opa1, and
- (iv) a fusion protein containing COX1, said COX1 homologue or said COX1 fragment;
- and
- (b) said second protein is selected from the group consisting of
- (1) THR S14 and Opa1,
- (2) a homologue of a protein selected from the group consisting of THR S14 and Opa1 having an amino acid sequence at least 90% identical to that of said protein and capable of interacting with COX1,
- (3) a fragment of a protein selected from the group consisting of THR S14 and Opa1 capable of interacting with COX1, and
- (4) a fusion protein containing a protein selected from the group consisting of THR S14 and Opa1, said protein homologue or said protein fragment, said method comprising:
- contacting said first protein with said second protein in the presence of a test compound and detecting the interaction between said first protein and said second protein;
- and
- contacting said first protein with said second protein in the absence of said test compound and detecting the interaction between said first protein and said second protein.
A composition comprising:
- a first expression vector having a nucleic acid encoding a first protein;
- and
- a second expression vector having a nucleic acid encoding a second protein, wherein:
- (a) said first protein is selected from the group consisting of
- (i) COX1,
- (ii) a COX1 homologue having an amino acid sequence at least 90% identical to that of COX1 and capable of interacting with a protein selected from the group consisting of THR S14 and Opa1,
- (iii) a COX1 fragment capable of interacting with a protein selected from the group consisting of THR S14 and Opa1, and
- (iv) a fusion protein containing COX1, said COX1 homologue or said COX1 fragment;
- and
- (b) said second protein is selected from the group consisting of
- (1) THR S14 and Opa1,
- (2) a homologue of a protein selected from the group consisting of THR S14 and Opa1 having an amino acid sequence at least 90% identical to that of said protein and capable of interacting with COX1,
- (3) a fragment of a protein selected from the group consisting of THR S14 and Opa1 capable of interacting with COX1, and
- (4) a fusion protein containing a protein selected from the group consisting of THR S14 and Opa1, said protein homologue or said protein fragment.
An expression vector comprising:
- (a) a first nucleic acid encoding a first protein selected from the group consisting of
- (i) COX1,
- (ii) a COX1 homologue having an amino acid sequence at least 90% identical to that of COX1 and capable of interacting with a protein selected from the group consisting of THR S14 and Opa1,
- (iii) a COX1 fragment capable of interacting with a protein selected from the group consisting of THR S14 and Opa1, and
- (iv) a fusion protein containing COX1, said COX1 homologue or said COX1 fragment;
- and
- (b) a second nucleic acid encoding a second protein selected from the group consisting of
- (1) THR S14 and Opa1,
- (2) a homologue of a protein selected from the group consisting of THR S14 and Opa1 having an amino acid sequence at least 90% identical to that of said protein and capable of interacting with COX1,
- (3) a fragment of a protein selected from the group consisting of THR S14 and Opa1 and capable of interacting with COX1, and
- (4) a fusion protein containing a protein selected from the group consisting of THR S14 and Opa1, said protein homologue or said protein fragment.
A host cell comprising:
- a first chimeric expression cassette having a nucleic acid encoding a first protein;
- and
- a second chimeric expression cassette having a nucleic acid encoding a second protein, wherein:
- (a) said first protein is selected from the group consisting of
- (i) COX1,
- (ii) a COX1 homologue having an amino acid sequence at least 90% identical to that of COX1 and capable of interacting with a protein selected from the group consisting of THR S14 and Opa1,
- (iii) a COX1 fragment capable of interacting with a protein selected from the group consisting of THR S14 and Opa1, and
- (iv) a fusion protein containing COX1, said COX1 homologue or said COX1 fragment;
- and
- (b) said second protein is selected from the group consisting of
- (1) THR S14 and Opa1,
- (2) a homologue of a protein selected from the group consisting of THR S14 and Opa1 and having an amino acid sequence at least 90% identical to that of said protein and capable of interacting with COX1,
- (3) a fragment of a protein selected from the group consisting of THR S14 and Opa1 and capable of interacting with COX1, and
- (4) a fusion protein containing a protein selected from the group consisting of THR S14 and Opa1, said protein homologue or said protein fragment.
Dependencies are inferred from the claim text.