Methods and materials for characterizing and modulating interaction between heregulin and HER3
AssigneeThe Regents of the University of California
InventorsElizabeth Singer, Ralf Landgraf, Dennis J. Slamon, David Eisenberg
Life of this patent
Estimating the term…
Abstract
The disclosure provided herein identifies and characterizes the domain in HER3 receptor that interacts with heregulin ligand. Typical embodiments of the invention disclosed herein include HER3 variant polypeptides having amino acid sequences which differ from the native HER3 polypeptide sequence and which have altered affinities for heregulin. Also disclosed herein are methods and materials for identifying compounds that bind to the heregulin binding domain in HER3 as well as methods and materials for modulating the interaction between HER3 and heregulin.
Claims25
A non-naturally occurring HER3 variant polypeptide consisting essentially of amino acid 20 to amino acid 329 of HER3 polypeptide sequence of SEQ ID NO:
- 2;
- wherein the HER3 variant polypeptide:
- (a) specifically binds to the heregulin polypeptide of SEQ ID NO:
- 4;
- and
- (b) exhibits an impaired ability to interact with the HER2 polypeptide of SEQ ID NO:
- 6.
A non-naturally occurring HER3 variant polypeptide consisting essentially of amino acid 19 to amino acid 329 of HER3 polypeptide sequence of SEQ ID NO:
- 2;
- wherein the HER3 variant polypeptide has the ability to inhibit the interaction between wild type HER3 having the polypeptide sequence of SEQ ID NO:
- 2 and heregulin having the polypeptide sequence of SEQ ID NO:
- 4.
A HER3 variant polypeptide comprising an amino acid sequence which differs from the native sequence HER3 polypeptide sequence of SEQ ID NO:
- 2 and has one or more of amino acid substitutions at the following residue position(s) in SEQ ID NO:
- 2:
- E43;
- N44;
- K51;
- E64;
- V66;
- and V110.
A method of identifying a compound which specifically binds the heregulin binding domain in a HER3 variant polypeptide comprising amino acid 20 to amino acid 329 of HER3 polypeptide sequence of SEQ ID NO:
- 2 comprising contacting the HER3 variant polypeptide with a test compound under conditions favorable to binding and determining whether the test compound specifically binds to the HER3 variant polypeptide such that a compound which binds to the HER3 variant polypeptide can be identified.
A method of determining whether a test compound modulates the interaction between heregulin polypeptide of SEQ ID NO:
- 4 and a HER3 variant polypeptide comprising amino acid 20 to amino acid 329 of HER3 polypeptide sequence of SEQ ID NO:
- 2 comprising the steps of:
- (a) contacting the HER3 variant polypeptide with a test compound under conditions favorable to binding;
- (b) contacting the HER3 variant polypeptide of step (a) with heregulin under conditions favorable to binding;
- (c) comparing the binding interaction between the HER3 variant polypeptide and heregulin of step (b) with the binding interaction between HER3 variant polypeptide and heregulin in the absence of the test compound such that a compound which modulates the interaction between heregulin and the HER3 variant polypeptide can be identified.
A method of inhibiting the interaction between a heregulin polypeptide having the sequence shown in SEQ ID NO:
- 4 and HER3 polypeptide having the sequence shown in SEQ ID NO:
- 2 comprising exposing the heregulin polypeptide to a non-naturally occurring HER3 variant polypeptide comprising amino acid 20 to amino acid 329 of HER3 polypeptide sequence of SEQ ID NO:
- 2;
- wherein the HER3 variant polypeptide specifically binds to the heregulin polypeptide of SEQ ID NO:
- 4;
- and exhibits an impaired ability to interact with the HER2 polypeptide of SEQ ID NO:
- 6.
Dependencies are inferred from the claim text.