Phage displayed PDZ domain ligands
AssigneeGenentech, Inc.
InventorsHeike A. Held, Laurence A. Lasky, Richard P. Laura, Sachdev S. Sidhu, Wai Lee Tan Wong, Yan Wu
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Abstract
The invention pertains to a method of identifying PDZ interacting polypeptides, said polypeptides, and uses of said polypeptides.
Claims50
A fusion protein comprising at least a portion of a phage coat protein bonded through the carboxyl-terminus thereof, optionally through a peptide linker, to a PDZ domain binding peptide, where the peptide contains 3-20 amino acid residues.
An assay for a PDZ domain binding compound, comprising:
- contacting a PDZ domain containing polypeptide with a candidate PDZ domain binding compound, and detecting binding of the polypeptide and compound.
An isolated polypeptide comprising a carboxy terminal amino acid sequence having the sequence of a member selected from the group consisting of SEQ ID NOs:1-181, 209-213, 241-601 and 709-714.
A method of inhibiting a polypeptide-polypeptide interaction, comprising:
- contacting a mixture comprising a first and a second polypeptide with an inhibitor of interaction between a PDZ domain and its ligand, wherein the first polypeptide comprises said PDZ domain and the second polypeptide comprises said ligand.
A method of screening for a substance that modulates interaction between a PDZ domain polypeptide and a molecule known to bind to the PDZ domain of said polypeptide comprising:
- (a) contacting a sample containing said polypeptide and molecule with a candidate substance;
- (b) determining amount of binding of said molecule to said polypeptide in the presence of said candidate substance;
- (c) comparing the amount of binding of step (b) with amount of binding of said molecule to said polypeptide under similar conditions in the absence of said candidate substance;
- whereby a difference in amount of binding as determined in (c) indicates that said candidate substance is a substance that modulates said interaction.
A method of screening for a substance that inhibits binding of a PDZ domain polypeptide to a molecule known to bind to the PDZ domain of said polypeptide comprising:
- (a) contacting a sample containing said polypeptide and molecule with a candidate substance;
- (b) determining amount of binding said molecule to said polypeptide in the presence of the candidate substance;
- (c) comparing the amount of binding of step (b) with amount of binding of said molecule to said polypeptide under similar conditions in the absence of the candidate substance;
- whereby a decrease in amount of binding of the polypeptide and said molecule in the presence of the candidate substance compared to the amount of binding in the absence of said candidate substance as determined in (c) indicates that said candidate substance is a substance that inhibits binding of the PDZ domain polypeptide to the molecule known to bind to the PDZ domain of said polypeptide.
A method of screening for a substance that increases binding of a PDZ domain polypeptide to a molecule known to bind to the PDZ domain of said polypeptide comprising:
- (a) contacting a sample containing said polypeptide and molecule with a candidate substance;
- (b) determining amount of binding said molecule to said polypeptide in the presence of the candidate substance;
- (c) comparing the amount of binding of step (b) with amount of binding of said molecule to said polypeptide under similar conditions in the absence of the candidate substance;
- whereby an increase in amount of binding of the polypeptide and said molecule in the presence of the candidate substance compared to the amount of binding in the absence of said candidate substance as determined in (c) indicates that said candidate substance is a substance that increases binding of the PDZ domain polypeptide to the molecule known to bind to the PDZ domain of said polypeptide.
A method comprising administering a substance to a subject with a condition associated with abnormal binding interaction of a PDZ domain polypeptide and a ligand, wherein said substance is a modulator of said binding interaction.
Dependencies are inferred from the claim text.