Functional antagonists of hedgehog activity
AssigneeCuris, Inc.
InventorsKevin Williams, Paul Rayhorn, Ellen A. Garber, R. Blake Pepinsky
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Abstract
Variants of hedgehog protein that contain N-terminal modifications are described that can block hedgehog function; thus allowing these variants to serve as functional antagonists. These peptides have a primary amino acid sequence lacking the ability to elicit a hedgehog-dependent response in C3H 10T1/2 cells but having the ability to bind to the hedgehog receptor, patched-1. Methods for producing such functional antagonists and methods of using the functional antagonists are also described.
Claims46
An isolated, functional antagonist of a hedgehog polypeptide, the antagonist characterized in that the antagonist can bind to a hedgehog receptor, but does not induce a hedgehog-dependent signaling response.
An isolated, Sonic hedgehog antagonist of Sonic, Desert, or Indian hedgehog polypeptide, wherein the antagonist comprises a Sonic hedgehog polypeptide characterized in that the antagonist can bind to a Sonic, Desert, or Indian hedgehog-responsive receptor, but cannot induce a Sonic, Desert, or Indian hedgehog-dependent signaling response.
An isolated, Desert hedgehog antagonist of Desert, Sonic, or Indian hedgehog polypeptide, wherein the antagonist comprises a Desert hedgehog polypeptide characterized in that the antagonist can bind to a Desert, Sonic, or Indian hedgehog-responsive receptor, but cannot induce a Sonic, Indian or Desert hedgehog-dependent signaling response.
An isolated, Indian hedgehog antagonist of Indian, Desert, or Sonic hedgehog polypeptide, wherein the antagonist comprises an Indian hedgehog polypeptide characterized in that the antagonist can bind to an Indian, Desert ,or Sonic hedgehog-responsive receptor, but cannot induce an Indian, Desert, or Sonic hedgehog-dependent signaling response.
An isolated, functional antagonist of a hedgehog polypeptide, selected from the group of polypeptides consisting of SEQ ID) NOS:
- 5, 6, 7, 8, 9 and 11.
An isolated, functional antagonist of a hedgehog polypeptide, the antagonist having the following properties:
- (i) the isolated antagonist binds a hedgehog receptor;
- and
- (ii) the isolated antagonist blocks alkaline phosphatase (AP) induction by mature hedgehog protein when tested in an AP assay.
A DNA sequence encoding a functional antagonist of a hedgehog polypeptide, the DNA sequence comprising a polynucleotide selected from the group consisting of:
- (a) SEQ ID NOS.:
- 16 and 17;
- (b) a polynucleotide that hybridizes to any of the foregoing sequences under standard hybridization conditions and that encodes a protein having the activity of a functional antagonist of a hedgehog polypeptide;
- and
- (c) a polynucleotide that codes on expression for a protein encoded by any of the foregoing polynucleotide sequences.
A method of making a functional hedgehog antagonist comprising altering an N-terminal cysteine residue of a mature hedgehog polypeptide from a residue corresponding to residue Cys-1 of mature Sonic hedgehog to a residue that does not correspond to residue Cys-1 of mature Sonic hedgehog.
A method of making a functional hedgehog antagonist protein comprising modifying a hedgehog polypeptide at an N-terminal cysteine residue of the hedgehog polypeptide.
A method of making a functional hedgehog antagonist protein comprising modifying a hedgehog polypeptide at an amino acid residue that is other than an N-terminal cysteine residue of the hedgehog polypeptide.
Dependencies are inferred from the claim text.