Receptor proteins

• US 20040142417A1
• Filed: 06/23/2003
• Published: 07/22/2004
• Est. Priority Date: 09/17/1993
• Status: Active Grant

• Current Assignee: Genetics Institute LLC (Pfizer Inc.)
• Original Assignee: Genetics Institute LLC (Pfizer Inc.)
• Inventors: Yamaji, Noboru, Thies, R. Scott, Wozney, John M., Celeste, Anthony J.
• Granted Patent: US 7,091,007 B2
• Time in Patent Office: Days
• US Class Current: 435/69.1
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: C07K 14/51 Bone morphogenetic factor; Osteogenins; Osteogenic factor; Bone-inducing factor
• CPC Class Codes: C07K 14/71 for growth factors; for growth regulators

Abstract

Novel serine/threonine receptor proteins and BMP receptor proteins are disclosed, as well as DNA molecules encoding said proteins and methods of using the receptor proteins. Further disclosed are truncated BMP receptor proteins and molecules which act as ligands to said BMP receptor proteins.

Claims

1. An isolated DNA molecule comprising a DNA sequence encoding a BMP receptor protein.

2. The DNA molecule of claim 1, wherein said DNA sequence is selected from the group consisting of:

(a) nucleotides 61 to 1656 of SEQ ID NO;

1;

(b) nucleotides 247 to 1752 of SEQ ID NO;

3; and

(c) sequences which hybridize to (a) or (b) under stringent hybridization conditions and encode a BMP receptor protein with the ability to bind to BMP in a binding assay.

4. The DNA molecule of claim 1, wherein said DNA sequence is selected from the group consisting of:

(a) nucleotides encoding for amino acids 24 to 532 of SEQ ID NO;

2; and

(b) nucleotides encoding for amino acids 8 to 502 of SEQ ID NO;

4; and

(c) sequences which hybridize to any of (a) or (b) under stringent hybridization conditions and encode a BMP receptor protein with the ability to bind to BMP in a binding assay.

6. A host cell transformed with the DNA molecule of claim 1.

7. A host cell transformed with the DNA molecule of claim 2.

8. A host cell transformed with the DNA molecule of claim 4.

3. An isolated DNA molecule comprising a DNA sequence encoding a serine/threonine kinase receptor protein, wherein the DNA sequence is selected from the group consisting of:

(a) nucleotides 474 to 2000 of SEQ ID NO;

5;

(b) nucleotides 80 to 1594 of SEQ ID NO;

7;

(c) nucleotides 83 to 1591 of SEQ ID NO;

9;

(d) sequences which hybridize to any of (c) to (e) under stringent hybridization conditions and encode a serine/threonine receptor protein.

5. An isolated DNA molecule comprising a DNA sequence encoding a serine/kinase receptor protein, wherein the DNA sequence is selected from the group consisting of:

(a) nucleotides encoding for amino acids 18 to 509 of SEQ ID NO;

6; and

(b) nucleotides encoding for amino acids 24 to 505 of SEQ ID NO;

8; and

(c) nucleotides encoding for amino acids 30 to 503 of SEQ ID NO;

10; and

(d) sequences which hybridize to any of (a) to (c) under stringent hybridization conditions and encode a serine/threonine kinase receptor protein.

9. A host cell transformed with the DNA molecule of claim 5.

10. An isolated DNA molecule having a sequence encoding a truncated BMP receptor protein which is characterized by the ability to bind to BMP in a binding assay, said DNA molecule comprising a DNA sequence selected from the group consisting of:

(a) nucleotide 61 to 507 of SEQ ID NO;

1;

(b) nucleotides encoding amino acids 1 to 149 of SEQ ID NO;

2;

(c) nucleotide 247 to 618 of SEQ ID NO;

3;

(d) nucleotides encoding amino acids 8 to 124 of SEQ ID NO;

4; and

(e) naturally occurring allelic sequences and equivalent degenerative codon sequences of (a), (b), (c) or (d).

11. A host cell transformed with the DNA molecule of claim 10.

12. A vector comprising a DNA molecule of claim 10 in operative association with an expression control sequence therefor.

13. A method for producing a purified truncated BMP receptor protein, said method comprising the steps of:

(a) culturing in a culture medium a host cell transformed with a DNA sequence according to claim 10, comprising a nucleotide sequence encoding a truncated BMP receptor protein; and

(b) recovering and purifying said truncated BMP receptor protein from the culture medium.

19. A method for producing a truncated BMP receptor protein, said method comprising the steps of:

(a) culturing in a culture medium a host cell according to claim 11, comprising a truncated nucleotide sequence encoding the ligand binding domain of a BMP receptor protein; and

(b) recovering and purifying said BMP receptor protein from the culture medium.

45. A ligand for the truncated BMP receptor protein produced by the method of claim 13, said ligand being selected from the group consisting of an BMP-like monoclonal antibody, a small peptide BMP analogue, and a small organic molecule BMP analogue.

46. The ligand of claim 45, comprising an BMP-like monoclonal antibody.

47. The ligand of claim 45, comprising a small peptide BMP analogue.

48. The ligand of claim 45, comprising a small organic molecule BMP analogue.

14. A purified truncated BMP receptor protein comprising the amino acid sequence from amino acid 24 to amino acid 149 of SEQ ID NO:

2.

49. A pharmaceutical composition comprising a compound first identified for such use as a ligand for the truncated BMP receptor produced by the method of claim 14.

50. The composition of claim 49, comprising an BMP-like monoclonal antibody.

51. The composition of claim 49, comprising a small peptide BMP analogue.

52. The composition of claim 49, comprising a small organic molecule BMP analogue.

15. A purified truncated BMP receptor protein comprising the amino acid sequence from amino acid 8 to amino acid 124 of SEQ ID NO:

4.

16. A purified truncated BMP receptor protein produced by the steps of:

(a) culturing in a culture medium a cell transformed with a DNA comprising the nucleotide sequence from nucleotide 61 to 507 of SEQ ID NO;

1; and

(b) recovering and purifying from said culture medium a protein comprising the amino acid sequence from amino acid 24 to amino acid 149 of a sequence selected from the group consisting of SEQ ID NO;

2.

17. A purified truncated BMP receptor protein produced by the steps of:

(a) culturing in a culture medium a cell transformed with a DNA comprising the nucleotide sequence from nucleotide 247 to 618 of SEQ ID NO;

3; and

(b) recovering and purifying from said culture medium a protein comprising the amino acid sequence from amino acid 8 to amino acid 124 of a sequence selected from the group consisting of SEQ ID NO;

4.

18. A method for producing a truncated BMP receptor protein, said method comprising the steps of:

(a) culturing in a culture medium a host cell transformed with a DNA sequence encoding a truncated BMP receptor protein, comprising a truncated nucleotide sequence encoding the ligand binding domain of a BMP receptor protein; and

(b) recovering and purifying said BMP receptor protein from the culture medium.

20. An isolated DNA molecule having a sequence encoding a truncated serine/threonine kinase receptor protein, said DNA molecule comprising a DNA sequence selected from the group consisting of:

(a) nucleotide 474 to 836 of SEQ ID NO;

5;

(b) nucleotides encoding amino acids 1 to 121 of SEQ ID NO;

6;

(c) nucleotide 80 to 445 of SEQ ID NO;

7;

(d) nucleotides encoding amino acids 1 to 122 of SEQ ID NO;

8;

(e) nucleotide 83 to 445 of SEQ ID NO;

9;

(f) nucleotides encoding amino acids 1 to 121 of SEQ ID NO;

10; and

(e) naturally occurring allelic sequences and equivalent degenerative codon sequences of (a) through (f).

21. A host cell transformed with the DNA molecule of claim 20.

22. A vector comprising a DNA molecule of claim 20 in operative association with an expression control sequence therefor.

23. A method for producing a purified truncated serine/threonine kinase receptor protein, said method comprising the steps of:

(a) culturing in a culture medium a host cell transformed with a DNA sequence according to claim 20, comprising a nucleotide sequence encoding a truncated serine/theeonine kinase receptor protein; and

(b) recovering and purifying said truncated serine/threonine kinase receptor protein from the culture medium.

28. A method for producing a truncated serine/threonine kinase receptor protein, said method comprising the steps of:

(a) culturing in a culture medium a host cell according to claim 21, comprising a truncated nucleotide sequence encoding the ligand binding domain of a serine/threonine kinase receptor protein; and

(b) recovering and purifying said truncated serine/threonine kinase receptor protein from the culture medium.

41. A pharmaceutical composition comprising a compound first identified for such use as a ligand for the truncated BMP receptor of claim 20.

42. The composition of claim 41, comprising an BMP-like monoclonal antibody.

43. The composition of claim 41, comprising a small peptide BMP analogue.

44. The composition of claim 41, comprising a small organic molecule BMP analogue.

24. A purified truncated serine/threonine kinase receptor protein comprising the amino acid sequence from amino acid 1 to amino acid 121 of SEQ ID NO:

6.

25. A purified truncated serine/threonine kinase receptor protein comprising the amino acid sequence from amino acid 1 to amino acid 122 of SEQ ID NO:

8.

26. A purified truncated serine/threonine kinase receptor protein comprising the amino acid sequence from amino acid 1 to amino acid 121 of SEQ ID NO:

10.

27. A method for producing a truncated serine/threonine kinase receptor protein, said method comprising the steps of:

(a) culturing in a culture medium a host cell transformed with a DNA sequence encoding a truncated serine/threonine kinase receptor protein, comprising a truncated nucleotide sequence encoding the ligand binding domain of a serine/threonine kinase receptor protein; and

(b) recovering and purifying said truncated serine/threonine kinase receptor protein from the culture medium.

29. A DNA molecule encoding a BMP receptor protein isolated through a method comprising:

(a) preparing a DNA fragment encoding a ligand binding domain of a known BMP receptor protein as a probe;

(b) screening a genomic or cDNA library, using the DNA fragment of (a) as a probe;

(c) isolating a DNA molecule which hybridized to the probe identified in step (b); and

(d) cloning a DNA molecule encoding a BMP receptor protein from the DNA molecule of step (c).

30. The BMP receptor protein of claim 29, wherein the known BMP receptor protein is CFK1-23a or CFK1-43a.

31. An isolated DNA molecule comprising the DNA sequence of CFK1-23a.

32. An isolated DNA molecule comprising the DNA sequence of CFK1-43a.

33. An isolated DNA molecule comprising the DNA sequence of CFK1-10a.

34. An isolated DNA molecule comprising the DNA sequence of W-101.

35. An isolated DNA molecule comprising the DNA sequence of W-120.

36. A composition comprising cells transformed with a DNA molecule comprising a DNA sequence encoding one or more BMP receptor proteins.

37. A ligand which binds to a truncated BMP receptor protein, said ligand being selected from the group consisting of an BMP-like monoclonal antibody, a small peptide BMP analogue, and a small organic molecule BMP analogue.

38. The ligand of claim 37, comprising a BMP-like monoclonal antibody.

39. The ligand of claim 37, comprising a small peptide BMP analogue.

40. The ligand of claim 37, comprising a small organic molecule BMP analogue.