1. A method of operating a cordless telephone comprising a remote handset and a base unit matched to said remote handset, the method comprising:
the remote handset playing a digital audio bit stream comprising music;
the remote handset digitally summing a digitally synthesized ring tone indicating an incoming call to said base unit with the digital audio bit stream to allow a user of said cordless telephone to hear said cordless telephone ringing along with said music; and
the remote handset muting said playing of said digital audio bit stream by an action initiated by a user of said cordless telephone when said cordless telephone receives a telephone call;
wherein said digital audio bit stream is either (i) pre-loaded before said step of playing or (ii) played substantially real-time as it is received by said cordless telephone.
2. The method according to claim 1, wherein:
said digital audio bit stream is pre-loaded before said step of playing.
3. The method of integrating a digital bit stream music player in a cordless telephone according to claim 2, wherein:
said digital audio bit stream is played substantially real-time as it is received by said cordless telephone.
4. The method according to claim 1, further comprising:
downloading said digital audio bit stream to said remote handset from a remote bit stream audio source.
5. The method according to claim 4, further comprising:
storing said downloaded digital audio bit stream in a base unit of said cordless telephone.
6. The method according to claim 4, wherein:
said remote bit stream audio source is accessible by said remote handset via an Internet.
7. The method according to claim 4, further comprising:
storing said downloaded digital audio bit stream in said remote handset of said cordless telephone.
8. The method according to claim 7, wherein:
said downloaded digital audio bit stream is stored in Flash memory in said remote handset.
9. The method according to claim 4, wherein:
said digital audio bit stream is comprised in an MPEG format.
10. The method according to claim 9, wherein:
said MPEG format is an MP3 format.
11. A cordless telephone, comprising:
a remote handset; and
a base unit matched to said remote handset, wherein said remote handset comprises:
means for playing a digital audio bit stream comprising music;
means for digitally summing a digitally synthesized ring tone indicating an incoming call to said base unit with a digital audio bit stream to allow a user of said cordless telephone to hear said cordless telephone ringing along with said music; and
means for muting said playing of said digital audio bit stream by an action initiated by a user of said cordless telephone when said cordless telephone receives a telephone call;
wherein said cordless telephone further comprises a means for downloading digital bit stream music to said remote handset from a remote bit stream audio source.
12. The cordless telephone according to claim 11, further comprising:
means for storing said downloaded digital bit stream music in a base unit of said cordless telephone.
13. The cordless telephone according to claim 11, wherein:
said remote bit stream audio source is accessible by said remote handset via an Internet.
14. The cordless telephone according to claim 11, further comprising:
means for decompressing MPEG formatted music into digital music samples for digital to analog output.
15. The cordless telephone according to claim 11, further comprising:
means for storing said downloaded digital bit stream music in said remote handset of said cordless telephone.
16. The cordless telephone according to claim 15, wherein:
said means for storing stores said downloaded digital bit stream music in Flash memory in said remote handset.
17. A cordless telephone, comprising:
a remote handset; and
a base unit matched to said remote handset, wherein said remote handset comprises:
a digital audio bit stream player adapted to play a digital audio bit stream comprising music;
a summer adapted to digitally sum a digitally synthesized ring tone indicating an incoming call to said base unit with the digital audio bit stream to allow a user of said cordless telephone to hear said cordless telephone ringing along with said music;
a means for muting the playing of said digital audio bit stream by an action initiated by a user of said cordless telephone when said cordless telephone receives a telephone call, and
a memory adapted to store said digital audio bit stream, wherein said cordless telephone is adapted to pre-load said digital audio bit stream into said memory before said digital audio bit stream player plays said digital audio bit stream.
18. A cordless telephone, comprising:
a remote handset;
a base unit matched to said remote handset, wherein:
said remote handset comprises:
a digital audio bit stream player adapted to play a digital audio bit stream comprising music;
a summer adapted to digitally sum a digitally synthesized ring tone indicating an incoming call to said base unit with the digital audio bit stream to allow a user of said cordless telephone to hear said cordless telephone ringing along with said music; and
a means for muting the playing of said digital audio bit stream by an action initiated by a user of said cordless telephone when said cordless telephone receives a telephone call;
said cordless telephone is adapted to receive said digital audio bit stream, and
said digital audio bit stream player is adapted to play said digital audio bit stream substantially real-time as it is received by said cordless telephone.
The claims below are in addition to those above.
All refrences to claim(s) which appear below refer to the numbering after this setence.
1. 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.
2. The isolated antagonist of claim 1, wherein the antagonist is characterized in that it does not induce a hedgehog-dependent signaling response in an assay selected from the group of assays consisting of: an alkaline phosphatase induction assay in CH310T12 cells; a ptc-1 expression assay in C3H10T12 cells; a gli-1 expression assay in C3H10T12 cells; and a patched-1 protein induction assay in C3H10T12 cells
3. The isolated antagonist of claim 1, wherein the antagonist is a hedgehog polypeptide that has an N-terminal cysteine residue altered from a residue corresponding to residue Cys-1 of a mature hedgehog polypeptide to a residue that does not correspond to residue Cys-1 of the mature hedgehog polypeptide.
4. The isolated antagonist of claim 3, lacking an N-terminal cysteine corresponding to Cys-1 of the mature hedgehog polypeptide.
5. The isolated antagonist of claim 3, wherein the antagonist includes an N-terminal extension moiety having an element that corresponds to a Cys-1 of the mature hedgehog polypeptide.
6. The isolated antagonist of claim 3, wherein the antagonist includes an N-terminal extension moiety having an element that replaces a Cys-1 of the mature hedgehog polypeptide.
7. The isolated antagonist of claim 3, wherein the antagonist is missing no greater than about 12 amino acids beginning from an N-terminal cysteine corresponding to a Cys-1 of the mature hedgehog polypeptide.
8. The isolated antagonist of claim 3, wherein the antagonist has a mutation of the N-terminal cysteine to another amino acid residue.
9. The isolated antagonist of claim 8, wherein the another amino acid is a polar or charged amino acid residue.
10. The isolated antagonist of claims 8 or 9, wherein the another amino acid residue is a serine residue.
11. The isolated antagonist of claim 3, wherein the antagonist further includes a modification to a C-terminal end of the antagonist.
12. The antagonist of claim 11, wherein the antagonist is missing no greater than about 11 amino acid residues from the C-terminal end.
13. The antagonist of claim 11, wherein the modification comprises a polypeptide linked to the C-terminal end, the polypeptide having an amino acid sequence unrelated to the antagonist polypeptide.
14. The antagonist of claim 13, wherein the polypeptide linked to the antagonist is selected from the group consisting of a glutathione-S-transferase, a DNA binding domain, a polymerase activating domain, a histidine tag, and an inmunoglobulin or portion thereof.
15. The antagonist of claim 11, wherein the C-terminal modification comprises a hydrophobic moiety linked to the C-terminal end.
16. The isolated antagonist of claim 1, wherein the N-terminal cysteine is in a modified form and corresponds to Cys-1 of the mature hedgehog polypeptide.
17. The isolated antagonist of claim 15, wherein the N-terminal cysteine is in an oxidized form.
18. The isolated antagonist of claim 3, wherein the Cys-1 of the mature hedgehog polypeptide is selected from the group consisting of (a) Cys-1 of mature Sonic hedgehog polypeptide; (b) Cys-1 of mature Indian hedgehog polypeptide; and (c) Cys-1 of mature Desert hedgehog polypeptide.
19. 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.
20. The isolated antagonist of claim 19, wherein the Sonic hedgehog polypeptide has an N-terminal cysteine residue altered from a residue corresponding to residue Cys-1 of a mature Sonic hedgehog polypeptide to a residue that does not correspond to residue Cys-1 of the mature Sonic hedgehog polypeptide.
21. 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.
22. The isolated antagonist of claim 21, wherein the Desert hedgehog polypeptide has an N-terminal cysteine residue altered from a residue corresponding to residue Cys-1 of a mature Desert hedgehog polypeptide to a residue that does not correspond to residue Cys-1 of the mature Desert hedgehog polypeptide.
23. 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.
24. The isolated antagonist of claim 23, wherein the Indian hedgehog polypeptide has an N-terminal cysteine residue altered from a residue corresponding to residue Cys-1 of a mature Indian hedgehog polypeptide to a residue that does not correspond to residue Cys-1 of the mature Indian hedgehog polypeptide.
25. 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.
26. 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.
27. The isolated antagonist of claim 26, wherein the hedgehog receptor is patched-1.
28. The isolated antagonist of claim 26 having the additional property of being unable to induce ptc-1 and gli-1 expression.
29. The isolated antagonist of claim 26, comprising a hedgehog polypeptide lacking an N-terminal cysteine corresponding to Cys-1 of a mature Sonic hedgehog.
30. An isolated DNA sequence encoding, upon expression, the functional antagonist of claims 1, 19, 21 or 23.
31. The DNA sequence of claim 30, wherein the DNA sequence encodes a hedgehog polypeptide that is a member of the vertebrate hedgehog family.
32. 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.
33. The DNA sequence of claim 32, wherein the DNA sequence encodes a hedgehog polypeptide that is at least 60% homologous to a polypeptide of the group selected from polypeptide SEQ ID NOS: 1-9 and 11.
34. A vector comprising the DNA sequence of claim 30.
35. A host cell containing the vector of claim 34.
36. A functional antagonist of a hedgehog polypeptide encoded by the DNA sequence contained within the vector of claim 34.
37. A therapeutic composition comprising the polypeptide of claims 1, 19, 21, 23, or 26 in an acceptable carrier.
38. 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.
39. The method of claim 38, wherein the step of altering comprises exposing hedgehog polypeptide to proteolysis.
40. A method of making a functional hedgehog antagonist protein comprising expressing protein from the host cell of claim 35 and purifying the protein.
41. A method of making a functional hedgehog antagonist protein comprising modifying a hedgehog polypeptide at an N-terminal cysteine residue of the hedgehog polypeptide.
42. 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.
43. A method of inhibiting hedgehog-dependent signaling in a subject, comprising administering to the subject the composition of claim 37 in an amount sufficient to inhibit hedgehog-dependent signaling in the subject
44. A method of inhibiting hedgehog-depending signaling in a subject, comprising administering to the subject the vector of claim 34, in an amount sufficient to inhibit hedgehog-dependent signaling in the subject.
45. The isolated antagonist of claim 3, wherein a moiety is linked to an amino acid residue of the antagonist, the moiety selected from the group consisting of polyethylene glycol and dextran.
46. The antagonist of claim 1, wherein the hedgehog-dependent signaling response is measured in a C3H10T12 alkaline phosphatase assay.