Kinase anchor protein muteins, peptides thereof and related documents

• US 20030232420A1
• Filed: 05/01/2003
• Published: 12/18/2003
• Est. Priority Date: 05/03/2002
• Status: Active Grant

• Current Assignee: Sequenom Incorporated (Laboratory Corporation Of America Holdings), Regents of the University of California (University of California)
• Original Assignee: Regents of the University of California (University of California)
• Inventors: Braun, Andreas, Self, Christopher, Cantor, Charles R., Cook, Charles, Olson, Gary, Taylor, Susan, Burns-Hamuro, Lora, Kammerer, Stefan M.
• Granted Patent: US 7,432,342 B2
• Time in Patent Office: Days
• US Class Current: 435/194
• CPC Class Codes: A01K 2207/15 Humanized animals
• CPC Class Codes: A01K 2217/00 Genetically modified animals
• CPC Class Codes: A01K 2217/072 maintaining or altering function, i.e. knock in
• CPC Class Codes: A01K 2227/105 Murine
• CPC Class Codes: A01K 2267/03 Animal model, e.g. for test or diseases
• CPC Class Codes: A01K 67/0275 Genetically modified vertebrates, e.g. transgenic
• CPC Class Codes: C07K 14/47 from mammals

Abstract

A-kinase anchor protein (AKAPs) muteins, peptides thereof, and nucleic acids encoding the peptides are provided herein. Also provided are transgenic animals, cells comprising transgenes and various methods employing such peptides.

Claims

1. A polypeptide that is a mutein of a D-AKAP2 polypeptide, wherein the mutein exhibits modified binding to a regulatory subunit of PKA compared to a native D-AKAP2.

2. The polyeptide of claim 1, wherein the native D-AKAP2 comprises a sequence of amino acids set forth as SEQ ID NOs:

1 or 2.

5. The polypeptide of claim 1 that exhibits enhanced binding to PKA-RIα

subunits.

6. The polypeptide of claim 5 that exhibits normal or reduced binding to PKA-RIIα

subunits.

7. The polypeptide of claim 1 that exhibits enhanced binding to PKA-RIIα

subunits relative to PKA-RIα

subunits.

8. The polypeptide of claim 7 that exhibits normal or reduced binding to PKA-RIα

subunits.

9. The polypeptide of claim 1 that exhibits enhanced binding to both RIα

and RIIα

subunits.

10. The polypeptide of claim 9 that exhibits reduced binding to PKA-RIα

subunits.

11. The polypeptide of claim 10 that exhibits normal or increased binding to PKA-RIIα

subunits.

12. The polypeptide of claim 1 that exhibits reduced binding to PKA-RIIα

subunits.

13. The polypeptide of claim 12 that exhibits normal or increased binding to PKA-RIα

subunits.

14. The polypeptide of claim 1 that exhibits reduced binding to both RIα

and RIIα

subunits.

19. The peptide of claim 1, wherein the peptide exhibits a preferred or exclusive binding to PKA-RIα

subunits relative to PKA-RIIα

subunits;

or enhanced binding to both RIα and

RIIα

subunits.

20. The peptide of claim 19, wherein the peptide is selected from the group consisting of peptides that correspond to the substitution in SEQ ID NOs:

1 or 2 of one, two or more;

of Q at residue 9 with F, I, L, V, H, M, R, T, W or Y;

of L at residue 12 with F, W or Y;

of V at residue 21 with I, L or W; and

of M at residue 25 with F, I, L, T, V, W or Y.

21. The peptide of claim 19, wherein V at residue 21 is substituted with W.

22. The peptide of claim 21, further comprising the substitution of either one or both of Q at residue 9 with F, and of M at residue 25 with F.

23. The peptide of claim 1, wherein the peptide exhibits a preferred or exclusive binding to PKA-RIIα

subunits relative to PKA-RIα

subunits.

24. The peptide of claim 23, wherein the peptide is selected from the group consisting of peptides that correspond to the substitution in SEQ ID NO:

2 of one, two or more;

of L at residue 12 with A, C, or K;

of A at residue 13 with F, H, I, K, L, M or N;

of W at residue 14 with C;

of K at residue 15 with C;

of K at residue 18 with C;

of M at residue 19 with C;

of S at residue 22 with C; and

of D at residue 23 with C.

25. The peptide of claim 1, wherein the peptide exhibits preferred or exclusive binding to PKA-RIIα

subunits relative to PKA-RIα

subunits.

26. The peptide of claim 25, wherein the peptide is selected from the group consisting of peptides that correspond to the substitution in SEQ ID NO:

1 of one, two or more;

of A at residue 13 with F, H, I, L, M and S;

of W at residue 14 with C;

of K at residue 15 with C;

of K at residue 18 with C;

of M at residue 19 with C;

of S at residue 22 with C; and

of D at residue 23 with C.

35. The peptide comprising any one of the peptides of claim 1, wherein when the peptide comprises amino acid positions corresponding to 1-27 of SEQ ID NOs:

1 or 2, the peptide further comprises a range of one or more amino acids, up to all of the amino acids, selected from amino acids 1-622 and 650-662 of SEQ ID NOs;

64 or 65, wherein the one or more of the selected amino acids are contiguous with amino acid positions corresponding to 623-649 of SEQ ID NOs;

64 or 65.

53. A cell, comprising heterologous nucleic acid that encodes a mammalian D-AKAP2 variant protein or portion comprising any one of the peptide sequences of claim 1 at the corresponding region in the mammalian D-AKAP2 variant protein.

71. A D-AKAP2 mutein, or a polypeptide fragment thereof, comprising a peptide region corresponding to an A-Kinsase binding (AKB) domain set forth as amino acids 623-649 of SEQ ID NOs:

64 or 65, wherein the sequence of the AKB domain peptide region corresponds to any of the peptide sequences of claim 1.

72. The D-AKAP2 mutein of claim 71, having 28 or more amino acid residues, wherein the amino acid sequence of the D-AKAP2 mutein additional to AKB domain peptide region, is 1 to 635 amino acids in length and corresponds to a contiguous region from amino acids 1-622 and/or 650-662 of SEQ ID NO:

64.

73. The D-AKAP2 mutein of claim 71, wherein the mutein is 662 amino acids in length, and further comprises amino acids 1-622 and 650-662 of SEQ ID NO:

64.

74. An isolated nucleic acid molecule, comprising a sequence of nucleotides that encodes the D-AKAP2 mutein of claim 71.

75. A nucleic acid vector, comprising the nucleic acid molecule of claim 74.

76. A cell containing the nucleic acid vector of claim 75.

77. A method of producing a D-AKAP2 mutein by growing the cell of claim 76 under conditions whereby the D-AKAP2 mutein is expressed;

and

isolating the mutein.

78. The method of claim 77, wherein the cell is a mammalian cell, yeast cell, insect cell or bacterial cell.

79. The method of claim 78, wherein the mammalian cell is a human cell.

80. A D-AKAP2 mutein produced by the method of claim 78.

81. A method for identifying a molecule that modulates the biological activity of an D-AKAP2 protein, comprising:

(a) combining the candidate molecule with a cell comprising a nucleotide sequence encoding a D-AKAP2 mutein of claim 71 or portion thereof that retains a biological activity exhibited by a full length variant protein, operably linked to a promoter such that the nucleotide sequence is expressed as an D-AKAP2 mutein or portion thereof in the cell; and

(b) determining the effect of the molecule upon the biological activity of the D-AKAP2 mutein or portion thereof.

82. The method of claim 81, wherein the biological activity of the D-AKAP2 mutein or portion thereof is determined by examining signal transduction in the cell.

83. The method of claim 81, wherein the biological activity is the binding of D-AKAP2 protein or portion thereof to protein kinase A.

84. The method of claim 81, wherein the biological activity of the D-AKAP2 protein or portion thereof is determined by examining protein phosphorylation.

85. The polypepeptide or peptide of claim 1, wherein the peptide is 27 amino acids in length.

105. A method of disrupting the binding of an Ile/Val(646) isoform of D-AKAP2 corresponding to SEQ ID NOs:

64 and 65, or fragment thereof, to the RIα

subunit of PKA comprising contacting the RIα

subunit with a peptide of claim 1.

106. A method of treating a subject manifesting a disease or disorder of signal transduction wherein there is an increased mitochondrial localization of PKA, the method comprising administering a peptide of claim 1 or peptidomimetic thereof.

3. A polypeptide that is a mutein of a D-AKAP2 polypeptide, wherein the mutein exhibits modified binding to a regulatory subunit of PKA compared to a control.

4. The polypeptide of claim 3, wherein the control is a polypeptide that consists essentially of the sequence of amino acids set forth as SEQ ID NOs:

1 or 2.

15. A peptide comprising between 15 and 39 amino acid residues, wherein the peptide comprises amino acids 7-21 of SEQ ID NOs:

1 or 2.

16. The peptide of claim 15, wherein the peptide binds to a regulatory subunit of PKA.

17. The peptide of claim 16, wherein the peptide binds to RIIα

.

18. The peptide of claim 17, wherein the peptide comprises amino acids 11-24 of SEQ ID NOs:

1 or 2 and binds to RIα

subunits.

36. The peptide of claim 17, wherein the peptide is selected from the group consisting of SEQ ID NOs:

4, 8, 9, 76, 77 and 86-88.

37. The peptide of claim 18, wherein the peptide is selected from the group consisting of SEQ ID NOs:

5, 6, 7, 66-75 and 83-85.

54. A method of disrupting the binding of an Ile/Val(646) isoform of D-AKAP2 corresponding to SEQ ID NOs:

64 and 65, or fragment thereof, to the RIα

subunit of PKA comprising contacting the RIα

subunit with a peptide of claim 15.

61. A method of treating a subject manifesting a disease or disorder of signal transduction wherein there is an increased mitochondrial localization of PKA, the method comprising administering a peptide of claim 15 or peptidomimetic thereof.

27. A peptide having substantially no ability to bind to PKA-RIα

subunit while maintaining the ability to bind to PKA-RIIα

subunit, compared to the peptide of SEQ ID NOs;

1 or 2, wherein the peptide is selected from the group consisting of;

28. A peptide having substantially no ability to bind to a PKA-RIα

subunit while maintaining a reduced ability to bind to a PKA-RIIα

subunit, compared to the peptide of SEQ ID NOs;

1 or 2, wherein the peptide is selected from the group consisting of;

29. A peptide that has enhanced binding to both PKA-RIα

subunit and PKA RIIα

subunit, compared to the peptide of SEQ ID NOs;

1 or 2, wherein the peptide is selected from the group consisting of;

30. A peptide that has enhanced ability to bind to PKA-RIα

subunit, while maintaining a normal or reduced ability to bind to PKA-RIIα

subunit, compared to the peptide of SEQ ID NOs;

1 or 2, wherein the peptide is selected from the group consisting of;

31. A peptide that has an ability to bind to PKA-RIα

subunit but substantially no ability to bind to PKA-RIIα

subunit, compared to the peptide of SEQ ID NOs;

1 or 2, wherein the peptide comprises

32. A peptide that has an enhanced ability to bind to PKA-RIα

subunit, and a reduced ability to bind to PKA-RIIα

subunit, compared to the peptide of SEQ ID NOs;

1 or 2, wherein the peptide has a D-amino acid at the position indicated in bold and wherein the peptide is selected from the group consisting of;

33. A peptide that has enhanced ability to bind to PKA-RIα

subunit, and a reduced ability to bind to PKA-RIIα

subunit, compared to the peptide of SEQ ID NOs;

1 or 2, wherein the peptide is selected from the group consisting of;

34. A peptide that has enhanced ability to bind to PKA-RIα

subunit, while maintaining a normal ability to bind to PKA-RIIα

subunit, compared to the peptide of SEQ ID NOs;

1 or 2, wherein the peptide is;

38. A transgenic non-human knock-in animal comprising a gene encoding an animal homolog of human D-AKAP2, wherein the gene has been modified to encode one, two or more amino acid substitutions in a 27-mer PKA binding region of the animal homolog of human D-AKAP2 corresponding to amino acids 623-649 of human D-AKAP2 set forth in SEQ ID NOs:

63, 64 or 65.

39. The transgenic animal of claim 38, wherein the amino acid substitution in the animal homolog of human D-AKAP2 is selected from the group consisting of substitutions that correspond to the substitution in SEQ ID NOs:

63, 64 or 65 of one, two or more;

of Q at residue 631 with F, I, L, V, H, M, R, T, W or Y;

of L at residue 634 with F, W or Y;

of V at residue 643 with I, L or W; and

of M at residue 647 with F, I, L, T, V, W or Y.

40. The transgenic animal of claim 39, wherein the modified animal homolog of human D-AKAP2 exhibits a preferred or exclusive binding to PKA-RIα

subunits relative to PKA-RIIα

subunits, or enhanced binding to both RIα and

RIIα

subunits, compared to the unmodified animal homolog of human D-AKAP2.

41. The transgenic animal of claim 38, wherein the amino acid substitution in the animal homolog of human D-AKAP2 is selected from the group consisting of substitutions that correspond to the substitution in SEQ ID NOs:

63, 64 or 65 of one, two or more;

of L at residue 634 with A, C, or K;

of A at residue 635 with F, H, I, K, L, M, N or S;

of W at residue 636 with C;

of K at residue 637 with C;

of K at residue 640 with C;

of M at residue 641 with C;

of S at residue 644 with C; and

of D at residue 645 with C.

42. The transgenic animal of claim 41, wherein the modified animal homolog of human D-AKAP2 exhibits a preferred or exclusive binding to PKA-RIIα

subunits relative to PKA-RIα

subunits, compared to the native unmodified animal homolog of human D-AKAP2.

43. The transgenic animal of claim 38, wherein the animal is a mouse, and wherein the amino acid substitution in the mouse D-AKAP2 is selected from the group consisting of substitutions that correspond to the substitution in SEQ ID NO:

56 of one, two or more;

of Q at residue 341 with F, I, L, V, H, M, R, T, W or Y;

of L at residue 344 with F, W or Y;

of V at residue 353 with I, L or W; and

of M at residue 357 with F, I, L, T, V, W or Y.

44. The transgenic animal of claim 43, wherein the modified animal homolog of human D-AKAP2 is mouse D-AKAP2 that exhibits a preferred or exclusive binding to PKA-RIα

subunits relative to PKA-RIIα

subunits, or enhanced binding to both RIα and

RIIα

subunits, compared to the native unmodified mouse D-AKAP2.

45. The transgenic animal of claim 43, wherein the amino acid substitution in the mouse D-AKAP2 corresponds to a single amino acid substitution in SEQ ID NO:

56 of Q at residue 341 with 1, and wherein the modified mouse D-AKAP2 exhibits a enhanced binding to PKA-RIα

subunits, and normal binding to PKA-RIIα

subunits compared to the native unmodified mouse D-AKAP2.

46. The transgenic animal of claim 43, wherein the amino acid substitution in the mouse D-AKAP2 corresponds to a single amino acid substitution in SEQ ID NO:

56 of V at residue 353 with W, and wherein the modified mouse D-AKAP2 exhibits a normal binding to PKA-RIα

subunits, and disrupted binding to PKA-RIIα

subunits compared to the native unmodified mouse D-AKAP2.

47. The transgenic animal of claim 43, wherein the amino acid substitution in the mouse D-AKAP2 corresponds to a triple amino acid substitution in SEQ ID NO:

56 of Q at residue 341 with F, of V at residue 353 with W, and of M at residue 357 with F, and wherein the modified mouse D-AKAP2 exhibits increased binding to RIα and

decreased binding affinity for RIIα

.

48. The transgenic animal of claim 47, wherein the modified mouse D-AKAP2 exhibits approximately 10-fold increased binding to RIα

and approximately 220-fold decreased binding affinity for RIIα

.

49. The transgenic animal of claim 38, wherein the animal is a mouse, and wherein the modified animal homolog of human D-AKAP2 is mouse D-AKAP2 that exhibits a preferred or exclusive binding to PKA-RIIα

subunits relative to PKA-RIα

subunits, compared to the native unmodified mouse D-AKAP2; and

wherein the amino acid substitution in the mouse D-AKAP2 is selected from the group consisting of substitutions that correspond to the substitution in SEQ ID NO;

56 of one, two or more;

of L at residue 344 with A, C, or K;

of A at residue 345 with F, H, I, K, L, M, N or S;

of W at residue 346 with C;

of K at residue 347 with C;

of K at residue 350 with C;

of M at residue 351 with C;

of S at residue 354 with C; and

of D at residue 355 with C.

50. The transgenic animal of claim 49, wherein the amino acid substitution in the mouse D-AKAP2 corresponds to a single amino acid substitution in SEQ ID NO:

56 of A at residue 345 with L, and wherein the modified mouse D-AKAP2 exhibits normal binding to PKA-RIIα

subunits, and disrupted binding to PKA-RIα

subunits compared to the native unmodified mouse D-AKAP2.

51. A cell, comprising heterologous nucleic acid that encodes a mammalian D-AKAP2 variant protein or portion that exhibits a preferred or exclusive binding to PKA-RIα

subunits relative to PKA-RIIα

subunits;

or enhanced binding to both RIα and

RIIα

subunits, compared to the unmodified full length D-AKAP2 protein, wherein the D-AKAP2 variant protein or portion thereof comprises at least one single amino acid substitution in the mammalian D-AKAP2 selected from the group consisting of substitutions that correspond to the substitution in SEQ ID NO;

63, 64 or 65 of one, two or more;

of Q at residue 631 with F, I, L, V, H, M, R, T, W or Y;

of L at residue 634 with F, W or Y;

of V at residue 643 with I, L or W; and

of M at residue 647 with F, I, L, T, V, W or Y.

52. A cell, comprising heterologous nucleic acid that encodes a mammalian D-AKAP2 variant protein or portion that exhibits a preferred or exclusive binding to PKA-RIIα

subunits relative to PKA-RIα

subunits, compared to the unmodified full length D-AKAP2 protein, wherein the D-AKAP2 variant protein or portion thereof comprises at least one single amino acid substitution in the mammalian D-AKAP2 selected from the group consisting of substitutions that correspond to the substitution in SEQ ID NO;

63, 64 or 65 of one, two or more;

of L at residue 634 with A, C, or K;

of A at residue 635 with F, H, I, K, L, M, N or S;

of W at residue 636 with C;

of K at residue 637 with C;

of K at residue 640 with C;

of M at residue 641 with C;

of S at residue 644 with C; and

of D at residue 645 with C.

55. A method for modulating the amount of PKA-RIα

bound to D-AKAP2 in a cell comprising;

changing the effective intracellular concentration of the Val(646) isoform of D-AKAP2 corresponding to SEQ ID NO;

65 in the cell.

56. The method of claim 55, wherein the intracellular concentration of the Val(646) isoform is increased, thereby increasing the amount of PKA-RIα

bound to D-AKAP2.

57. The method of claim 55, wherein the intracellular concentration of the Val(646) isoform is decreased, thereby decreasing the amount of PKA-RIα

bound to D-AKAP2.

58. A method for altering the intracellular location of PKA in a cell comprising:

changing the effective intracellular concentration of the Val(646) isoform of D-AKAP2 corresponding to SEQ ID NO;

65 in the cell.

59. The method of claim 58, wherein the intracellular concentration of the Val(646) isoform is increased, thereby increasing the amount of PKA localized to the mitochondria.

60. The method of claim 58, wherein the intracellular concentration of the Val(646) isoform is decreased, thereby decreasing the amount of PKA localized to the mitochondria.

62. A method of increasing the longevity of a subject in need thereof, comprising identifying a subject having the 646(Val) isoform of D-AKAP2 therein, and treating the subject with an agent that disrupts the binding the 646(Val) isoform of D-AKAP2 to RIα

subunit of PKA.

63. The method of claim 62, wherein the agent is a mutein of a D-AKAP2 polypeptide, or peptidomimetic thereof.

64. A method for altering the ratio of PKA-RIα

/PKA-RIIα

bound to D-AKAP2 in a cell comprising;

increasing the concentration of a Val(646) variant of D-AKAP2 corresponding to SEQ ID NO;

65 in a cell, thereby increasing the ratio of PKA-RIα

PKA/PKA-RIIα

bound to D-AKAP2 in a cell.

65. A method of decreasing the concentration of cAMP required to stimulate a cAMP mediated signalling pathway, comprising:

increasing the concentration of a Val(646) variant of D-AKAP2 corresponding to SEQ ID NO;

65 in a cell.

66. A method of screening for agents that decrease the binding of a Val(646) variant of D-AKAP2 with RIα

PKA, comprising;

combining a candidate agent with a cell comprising a nucleotide sequence which encodes a Val(646) variant D-AKAP protein corresponding to SEQ ID NO;

65, operably linked to a promoter such that the nucleotide sequence is expressed as a D-AKAP2 protein in the cell; and

determining the effect of the agent upon the localization of PKA to the mitochondria, wherein a decrease in localization to the mitochondria identifies an agent that decreases the binding of a Val(646) variant of D-AKAP2 with RIα

PKA.

67. A method of screening for agents that decrease the binding of a Val(646) variant of D-AKAP2 to an RIα

subunit of PKA, comprising;

combining a candidate agent with an admixture comprising RIα and

a D-AKAP2 peptide sequence that binds to RIα

; and

determining the effect of the agent upon the localization of PKA to a mitochondria.

68. The method of claim 67, wherein the candidate agent is combined with the admixture in a cell-free system.

69. The method of claim 68, wherein the candidate agent is combined with the admixture intracellularly.

70. The method claim 68, wherein the peptide sequence is a mutein of a D-AKAP2 polypeptide, or peptidomimetic thereof.

86. A peptide, comprising between 12 and 39 amino acid residues, wherein the peptide comprises a segment of SEQ ID NOs:

1 or 2, wherein the length of the segment ranges from at least amino acids 12-23 of SEQ ID NOs;

1 or 2 up to amino acids 1-27 of SEQ ID NOs;

1 or 2.

87. The peptide of claim 86, wherein the peptide binds to a regulatory subunit of PKA.

88. The peptide of claim 87, wherein the peptide binds to RIIα

subunits.

89. The peptide of claim 88, wherein the peptide comprises amino acids 11-24 of SEQ ID NOs:

1 or 2 and binds to RIα

subunits.

90. The peptide of claim 87, wherein the peptide exhibits a preferred or exclusive binding to PKA-RIα

subunits relative to PKA-RIIα

subunits;

or enhanced binding to both RIα and

RIIα

subunits.

91. The peptide of claim 90, wherein the peptide is selected from the group consisting of peptides that correspond to the substitution in SEQ ID NOs:

1 or 2 of one, two or more;

of Q at residue 9 with F, I, L, V, H, M, R, T, W or Y;

of L at residue 12 with F, W or Y;

of V at residue 21 with I, L or W; and

of M at residue 25 with F, I, L, T, V, W or Y.

92. The peptide of claim 91, wherein V at residue 21 is substituted with W.

93. The peptide of claim 92, further comprising the substitution of either one or both of Q at residue 9 with F, and of M at residue 25 with F.

94. The peptide of claim 87, wherein the peptide exhibits a preferred or exclusive binding to PKA-RIIα

subunits relative to PKA-RIα

subunits.

95. The peptide of claim 94, wherein the peptide is selected from the group consisting of peptides that correspond to the substitution in SEQ ID NO:

2 of one, two or more;

of L at residue 12 with A, C, or K;

of A at residue 13 with F, H, I, K, L, M or N;

of W at residue 14 with C;

of K at residue 15 with C;

of K at residue 18 with C;

of M at residue 19 with C;

of S at residue 22 with C; and

of D at residue 23 with C.

96. The peptide of claim 87, wherein the peptide exhibits preferred or exclusive binding to PKA-RIIα

subunits relative to PKA-RIα

subunits.

97. The peptide of claim 96, wherein the peptide is selected from the group consisting of peptides that correspond to the substitution in SEQ ID NO:

1 of one, two or more;

of A at residue 13 with F, H, I, L, M and S;

of W at residue 14 with C;

of K at residue 15 with C;

of K at residue 18 with C;

of M at residue 19 with C;

of S at residue 22 with C; and

of D at residue 23 with C.

98. The peptide of claim 86, wherein when the peptide comprises amino acid positions corresponding to 1-27 of SEQ ID NO:

1 or 2, the peptide further comprises a range of one or more amino acids, up to all of the amino acids, selected from amino acids 1-622 and 650-662 of SEQ ID NOs;

64 or 65, wherein the one or more of the selected amino acids are contiguous with amino acid positions corresponding to 623-649 of SEQ ID NOs;

64 or 65.

99. A D-AKAP2 mutein, or a polypeptide fragment thereof, comprising a peptide region corresponding to an A-Kinsase binding (AKB) domain set forth as amino acids 623-649 of SEQ ID NOs:

64 or 65, wherein the sequence of the AKB domain peptide region corresponds to any of the peptide sequences of claim 86.

100. The D-AKAP2 mutein of claim 99, having 28 or more amino acid residues, wherein the amino acid sequence of the D-AKAP2 mutein additional to AKB domain peptide region, is 1 to 635 amino acids in length and corresponds to a contiguous region from amino acids 1-622 and/or 650-662 of SEQ ID NO:

64.

101. The D-AKAP2 mutein of claim 99, wherein the mutein is 662 amino acids in length, and further comprises amino acids 1-622 and 650-662 of SEQ ID NO:

64.

102. An isolated nucleic acid molecule, comprising a sequence of nucleotides that encodes the D-AKAP2 mutein of claim 99.

103. A nucleic acid vector, comprising the nucleic acid molecule of claim 102.

104. A peptide selected from SEQ ID NOs:

3-9.