Human gtp-rho binding protein 2

• US 20040068380A1
• Filed: 09/15/2003
• Published: 04/08/2004
• Est. Priority Date: 05/23/2001
• Status: Abandoned Application

• Current Assignee: Amersham PLC (General Electric Company)
• Original Assignee: Amersham PLC (General Electric Company)
• Inventors: Shannon, Mark E.
• Application Number:
  US10/663,470
• Publication Number:
  US 20040068380A1
• Time in Patent Office: Days
• US Class Current: 702/20
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: A01K 2217/075 inducing loss of function, i.e. knock out
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 2039/53 DNA (RNA) vaccination
• 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: C07K 14/47 from mammals
• CPC Class Codes: C07K 14/4748 Tumour specific antigens; Tumour rejection antigen precursors [TRAP], e.g. MAGE
• CPC Class Codes: C07K 14/475 Growth factors; Growth regulators
• CPC Class Codes: C07K 14/705 Receptors; Cell surface antigens; Cell surface determinants tumour specific antigens C07K14/4748
• CPC Class Codes: C07K 2319/00 Fusion polypeptide
• CPC Class Codes: C07K 2319/02 containing a signal sequence
• CPC Class Codes: C07K 2319/40 containing a tag for immunodetection, or an epitope for immunisation
• CPC Class Codes: C07K 2319/60 containing spectroscopic/fluorescent detection, e.g. green fluorescent protein [GFP]
• CPC Class Codes: C12N 15/1034 Isolating an individual clone by screening libraries
• CPC Class Codes: C12N 15/66 General methods for inserting a gene into a vector to form a recombinant vector using cleavage and ligation; Use of non-functional linkers or adaptors, e.g. linkers containing the sequence for a restriction endonuclease
• CPC Class Codes: C12Q 1/6809 Methods for determination or identification of nucleic acids involving differential detection
• CPC Class Codes: C12Q 1/6837 using probe arrays or probe chips C12Q1/6874 takes precedence
• CPC Class Codes: C12Q 1/6876 Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
• CPC Class Codes: C12Q 1/6883 for diseases caused by alterations of genetic material
• CPC Class Codes: C12Q 1/6886 for cancer immunoassay for cancer G01N33/574
• CPC Class Codes: C12Q 2539/105 : Involving introns, exons, or splice junctions
• CPC Class Codes: C12Q 2565/501 : being an array of oligonucleotides
• CPC Class Codes: C12Q 2600/136 : Screening for pharmacological compounds
• CPC Class Codes: C12Q 2600/156 : Polymorphic or mutational markers
• CPC Class Codes: C12Q 2600/158 : Expression markers
• CPC Class Codes: G16B 25/00 : ICT specially adapted for hybridisation; ICT specially adapted for gene or protein expression
• CPC Class Codes: G16B 30/00 : ICT specially adapted for sequence analysis involving nucleotides or amino acids
• CPC Class Codes: Y02A 90/10 : Information and communication technologies [ICT] supporting adaptation to climate change, e.g. for weather forecasting or climate simulation

Abstract

The invention provides isolated nucleic acids that encode human GRBP2, and fragments thereof, vectors for propagating and expressing human GRBP2 nucleic acids, host cells comprising the nucleic acids and vectors of the present invention, proteins, protein fragments, and protein fusions of the human GRBP2, and antibodies thereto. The invention further provides transgenic cells and non-human organisms comprising human GRBP2 nucleic acids, and transgenic cells and non-human organisms with targeted disruption of the endogenous orthologue of the human GRBP2 gene. The invention further provides pharmaceutical formulations of the nucleic acids, proteins, and antibodies of the present invention, and diagnostic, investigational, and therapeutic methods based on the human GRBP2 nucleic acids, proteins, and antibodies of the present invention.

Claims

1. An isolated nucleic acid comprising a nucleotide sequence selected from the group consisting of:

(i) the nucleotide sequence of SEQ ID NO;

1;

(ii) the nucleotide sequence of SEQ ID NO;

2;

(iii) a degenerate variant of the nucleotide sequence of SEQ ID NO;

2;

(iv) a nucleotide sequence that encodes a polypeptide having the sequence of SEQ ID NO;

3;

(v) a nucleotide sequence that encodes a polypeptide having the sequence of SEQ ID NO;

3 with conservative amino acid substitutions;

(vi) at least 17 contiguous nucleotides of SEQ ID NO;

4;

(vii) at least 17 contiguous nucleotides of SEQ ID NO;

6;

(viii) a degenerate variant of the nucleotide sequence of SEQ ID NO;

6;

(viii) a nucleotide sequence that encodes a polypeptide having the sequence of SEQ ID NO;

7;

(ix) the complement of any one of (i)-(viii).

2. The isolated nucleic acid of claim 1 wherein said nucleic acid, or the complement of said nucleic acid, encodes a polypeptide which interacts with Rho and/or PDZ domain-containing proteins.

3. The isolated nucleic acid of claim 1, wherein said nucleic acid, or the complement of said nucleic acid, is expressed in kidney, colon, adrenal, adult liver, bone marrow, brain, fetal liver, heart, hela, lung, placenta, prostate and skeletal muscle.

4. The isolated nucleic acid molecule of any one of claims 1-3, wherein said nucleic acid molecule is operably linked to one or more expression control elements.

5. A replicable vector comprising an isolated nucleic acid molecule of any one of claims 1-3.

6. A replicable vector comprising an isolated nucleic acid molecule of claim 4.

7. The isolated nucleic acid molecule of any of claims 1-3, attached to a substrate.

8. A host cell transformed to contain the nucleic acid molecule of any one of claims 1-3, or the progeny thereof.

9. A host cell transformed to contain the nucleic acid molecule of claim 4, or the progeny thereof.

10. A host cell transformed to contain the nucleic acid molecule of claim 5, or the progeny thereof.

11. A host cell transformed to contain the nucleic acid molecule of claim 6, or the progeny thereof.

12. A method for producing a polypeptide, the method comprising:

culturing the host cell of claim 8 under conditions in which the protein encoded by said nucleic acid molecule is expressed.

13. A method for producing a polypeptide, the method comprising:

culturing the host cell of claim 9 under conditions in which the protein encoded by said nucleic acid molecule is expressed.

14. A method for producing a polypeptide, the method comprising:

culturing the host cell of claim 10 under conditions in which the protein encoded by said nucleic acid molecule is expressed.

15. A method for producing a polypeptide, the method comprising:

culturing the host cell of claim 11 under conditions in which the protein encoded by said nucleic acid molecule is expressed.

16. An isolated polypeptide produced by the method of claim 12.

17. An isolated polypeptide produced by the method of claim 13.

18. An isolated polypeptide produced by the method of claim 14.

19. An isolated polypeptide produced by the method of claim 15.

24. A method of modulating the expression of a nucleic acid according to claim 1, the method comprising:

administering an effective amount of an agent which modulates the expression of a nucleic acid according to claim 1.

26. A transgenic non-human animal modified to contain a nucleic acid molecule of any one of claims 1-3.

27. A transgenic non-human animal modified to contain a nucleic acid molecule of claim 4.

28. A transgenic non-human animal modified to contain a nucleic acid molecule of claim 5.

29. A transgenic non-human animal modified to contain a nucleic acid molecule of claim 6.

33. A pharmaceutical composition comprising the nucleic acid of any one of claims 1-3 and a pharmaceutically acceptable excipient.

34. A pharmaceutical composition comprising the nucleic acid of claim 4 and a pharmaceutically acceptable excipient.

35. A pharmaceutical composition comprising the nucleic acid of claim 5 and a pharmaceutically acceptable excipient.

36. A pharmaceutical composition comprising the nucleic acid of claim 6 and a pharmaceuticallky acceptable excipient.

43. A method for treating or preventing a disorder associated with decreased expression or activity of human GRBP2, the method comprising administering to a subject in need of such treatment an effective amount of the pharmaceutical composition of claim 37 or 41, or a pharmaceutical composition comprising a nucleic acid of claim 1 in a pharmaceutically acceptable carrier.

45. A diagnostic composition comprising the nucleic acid of claim 1, said nucleic acid being detectably labeled.

49. The diagnostic composition of any one of claims 45-48, wherein said composition is further suitable for in vivo administration.

50. A microarray wherein at least one probe of said array is a nucleic acid according to any one of claims 1-3.

51. A method for detecting a target nucleic acid in a sample, said target being a nucleic acid of any one of claims 1-3, the method comprising:

a) hybridizing the sample with a probe comprising at least 30 contiguous nucleotides of a sequence complementary to said target nucleic acid in said sample under hybridization conditions sufficient to permit detectable binding of said probe to said target, and

b) detecting the presence or absence, and optionally the amount, of said binding.

20. An isolated polypeptide selected from the group consisting of:

(a) an isolated polypeptide comprising the amino acid sequence of SEQ ID NO;

3;

(b) an isolated polypeptide comprising a fragment of at least 6 amino acids of SEQ ID NO;

7; and

(c) an isolated polypeptide according to (a) or (b) in which at least 95% of deviations from the sequence of (a) or (b) are conservative substitutions.

22. A method of identifying binding partners for a polypeptide according to claim 20, the method comprising:

contacting said polypeptide to a potential binding partner; and

determining if the potential binding partner binds to said polypeptide.

23. The method of claim 22, wherein said contacting is performed in vivo.

25. A method of modulating at least one activity of a polypeptide according to claim 20, the method comprising:

administering an effective amount of an agent which modulates at least one activity of a polypeptide according to claim 20.

30. A transgenic non-human animal unable to express the endogenous orthologue of the protein of claim 20.

37. A pharmaceutical composition comprising the polypeptide of claim 20 and a pharmaceutically acceptable excipient.

39. A purified agonist of the polypeptide of claim 20.

40. A purified antagonist of the polypeptide of claim 20.

41. A pharmaceutical composition comprising the agonist of claim 39.

42. A pharmaceutical composition comprising the antagonist of claim 40.

46. A diagnostic composition comprising the polypeptide of claim 20, said polypeptide being detectably labeled.

52. A fusion protein, said fusion protein comprising a polypeptide of claim 20 fused to a heterologous amino acid sequence.

53. The fusion protein of claim 52, wherein said heterologous amino acid sequence is a detectable moiety.

54. The fusion protein of claim 53, wherein said detectable moiety is fluorescent.

55. The fusion protein of claim 52, wherein said heterologous amino acid sequence is an Ig Fc region.

21. An isolated antibody or antigen-binding fragment or derivative thereof, the binding of which can be competitively inhibited by an isolated polypeptide having the sequence of SEQ ID NO:

7, or at least 6 contiguous amino acids thereof.

38. A pharmaceutical composition comprising the antibody or antigen-binding fragment or derivative thereof of claim 21 and a pharmaceutically acceptable excipient.

44. A method for treating or preventing a disorder associated with increased expression or activity of human GRBP2, the method comprising administering to a subject in need of such treatment an effective amount of the pharmaceutical composition of claim 38 or 42.

47. A diagnostic composition comprising the antibody or antigen-binding fragment or derivative thereof of claim 21.

48. The diagnostic composition of claim 47, wherein said antibody or antigen-binding fragment or derivative thereof is detectably labeled.

31. A method of diagnosing a disease caused by mutation in human GRBP2, comprising:

detecting said mutation in a sample of nucleic acids that derives from a subject suspected to have said disease.

32. A method of diagnosing or monitoring a disease caused by altered expression of human GRBP2, comprising:

determining the level of expression of human GRBP2 in a sample of nucleic acids or proteins that derives from a subject suspected to have said disease, alterations from a normal level of expression providing diagnostic and/or monitoring information.

56. A method of screening for agents that modulate the expression of human GRBP2, the method comprising:

contacting a cell or tissue sample believed to express human GRBP2 with a chemical or biological agent, and then

comparing the amount of human GRBP2 expression with that of a control.