Claims
- 1. A molecule with two pendant phenylarsine moieties according to the general structural Formula (I) and tautomers, acids, and salts thereof:
- 2. The molecule according to claim 1, wherein R8 has a minimum length of about 3.5 Ångstroms and a maximum length of about 10 Ångstroms.
- 3. The molecule according to claim 1, wherein R8 has a minimum length of about 1 atom and a maximum length of about 9 atoms.
- 4. The molecule according to claim 1, wherein the detectable group is selected from the group consisting of a fluorescent moiety, a phosphorescent moiety, a luminescent moiety, an absorbent moiety, a photosensitizer, a spin label, a radioisotope, an isotope detectable by nuclear magnetic resonance, a paramagnetic atom, a heavy atom, a hapten, a crosslinking agent, a cleavage agent, and combinations thereof.
- 5. The molecule according to claim 1, wherein X is a fluorescent moiety.
- 6. The molecule according to claim 1, wherein X is derived from a cyanine dye.
- 7. The molecule according to claim 1, wherein X is derived from a squaraine dye.
- 8. The molecule according to claim 1, where X is selected from the group consisting of:
- 9. The molecule according to claim 1, wherein n is 1, 2 or 3.
- 10. The molecule according to claim 1, wherein X is selected from the group consisting of:
- 11. The molecule according to claim 10, wherein n′ is 0, 1, or 2.
- 12. The molecule according to claim 1, wherein said molecule is capable of traversing a biological membrane.
- 13. A molecule with two pendant phenylarsine moieties according to the general structural Formula (VI):
- 14. A molecule with two pendant phenylarsine moieties according to the general structural Formula (VII):
- 15. A molecule with two pendant phenylarsine moieties according to the general structural Formula (VIII):
- 16. A molecule with two pendant phenylarsine moieties according to the general structural Formula (IX):
- 17. A molecule with two pendant phenylarsine moieties according to the general structural Formula (X):
- 18. A molecule with two pendant phenylarsine moieties according to the general structural Formula (XI):
- 19. A labeled target material comprising:
a target sequence including an amino acid sequence of the form C(X)iC(X)jC(X)kC, wherein C is cysteine, X is any amino acid, and i, j, and k are each independently 0 or an integer of from 1 to 6, said target sequence being bonded with a molecule according to Formula (I).
- 20. The target material according to claim 19, wherein said target material is a polypeptide.
- 21. The target material according to claim 19, wherein said target sequence is CC(X)jCC.
- 22. The target material according to claim 21, wherein said target sequence is selected from the group consisting of SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, and SEQ ID NO:4.
- 23. The target material according to claim 21 wherein said target sequence is selected from group consisting of SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8.
- 24. The target material according to claim 19, wherein said target sequence is CXXC(X)jCXXC.
- 25. The target material according to claim 24 wherein said target sequence is selected from the group consisting of SEQ ID NO: 9, SEQ ID NO: 10 and SEQ ID NO: 11.
- 26. A detectable complex comprising:
a molecule according to Formula (I); and a target sequence bonded with said molecule, said target sequence including an amino acid sequence of the form: C(X)iC(X)jC(X)kC, wherein C is cysteine, X is any amino acid, and i, j, and k are each independently 0 or an integer of from 1 to 6.
- 27. The complex according to claim 26, wherein said target material is a polypeptide.
- 28. The complex according to claim 26, wherein said target sequence is CC(X)jCC.
- 29. The complex according to claim 28, wherein said target sequence is selected from the group consisting of SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3, and SEQ ID NO:4.
- 30. The complex according to claim 28, wherein said target sequence is selected from the group consisting of SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO: 8.
- 31. The complex according to claim 26, wherein said target sequence is CXXC(X)jCXXC.
- 32. The complex according to claim 31, wherein said target sequence is selected from the group consisting of SEQ ID NO: 9, SEQ ID NO: 10 and SEQ ID NO: 11.
- 33. A method for imparting detectable properties to at least one target material of interest, the method comprising the step of reacting:
(a) a target material having a target sequence of the form: C(X)iC(X)jC(X)kC, wherein C is cysteine, X is any amino acid, and i, j, and k are each independently 0 or an integer of from 1 to 6, with (b) a molecule according to Formula (I), under conditions sufficient to permit bis-phenylarsine moieties of said molecule to bond to said target sequence.
- 34. The method according to claim 33, wherein said target material is a polypeptide.
- 35. The method according to claim 33, wherein said target sequence is CC(X)jCC.
- 36. The method according to claim 35, wherein said target sequence is selected from the group consisting of SEQ ID NO: 1, SEQ ID NO:2, SEQ ID NO:3 and SEQ ID NO:4.
- 37. The method according to claim 35 wherein said target sequence is selected from the group consisting of SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7 and SEQ ID NO:8.
- 38. The method according to claim 33, wherein said target sequence is CXXC(X)jCXXC.
- 39. The method according to claim 38, wherein said target sequence is selected from the group consisting of SEQ ID NO: 9, SEQ ID NO: 10 and SEQ ID NO: 11.
- 40. A method for detecting at least one target material of interest, said method comprising the steps of:
(a) providing a target material containing a target sequence of the form: C(X)iC(X)jC(X)kC, wherein C is cysteine, X is any amino acid, and i, j, and k are each independently 0 or an integer of from 1 to 6; (b) incubating said target material with a molecule according to Formula (I) having a detectable group, for a time period sufficient to allow labeling of said target material; and (c) detecting said detectable group, thereby detecting said target material.
- 41. The method according to claim 40, wherein said target material is located within a material selected from the group consisting of a cuvette, a microtiter plate, a capillary, a flow cell, a test tube, a gel, a blot, and a biological sample.
- 42. The method according to claim 40, wherein step (b) or (c) is performed in a gel matrix.
- 43. The method according to claim 40, wherein step (b) or (c) is performed in a complex mixture of components.
- 44. The method according to claim 40, wherein labeled target material is separated from other components following step (b).
- 45. The method according to claim 40, wherein labeled target material is not separated from other components following step (b).
- 46. The method according to claim 40, wherein said target material is a polypeptide.
- 47. The method according to claim 40, wherein said detectable group is a fluorescent moiety.
- 48. The method according to claim 40, wherein said step (c) includes detecting a fluorescence property.
- 49. The method according to claim 48, wherein said fluorescence property is at least one of a fluorescence-emission intensity, a fluorescence lifetime, a fluorescence polarization, a fluorescence anisotropy, and a fluorescence correlation.
- 50. A method for determining the localization, concentration or interactions of at least one target material of interest on or within a cell, tissue, organ or organism, said method comprising the steps of:
(a) providing a cell, tissue, organ, or organism containing a target material containing havinga target sequence of the form: C(X)iC(X)jC(X)kC, wherein C is cysteine, X is any amino acid, and i, j, and k are each independently 0 or an integer of from 1 to 6; (b) contacting said cell, tissue, organ, or organismwith a molecule according to Formula (I) having a detectable group, for a time period sufficient to allow labeling of said target material; and (c) detecting said detectable group, thereby determining the localization, concentration, or interactions of said target material.
- 51. The method according to claim 50, wherein said target material is a polypeptide.
- 52. The method according to claim 50, wherein said detectable group is a fluorescent moiety.
- 53. The method according to claim 50, wherein said step (c) includes detecting a fluorescence property.
- 54. The method according to claim 53, wherein said fluorescence property is at least one of a fluorescence-emission intensity, a fluorescence lifetime, a fluorescence polarization, a fluorescence anisotropy, and a fluorescence correlation.
- 55. An assay method for monitoring a binding process comprising:
(a) reacting a first component of a specific binding pair with a second component of said pair, said first component being labeled with a molecule according to Formula (I) having a detectable group; and (b) monitoring said reaction by monitoring a change in a signal of said detectable group.
- 56. An assay method for monitoring a binding process, comprising:
(a) reacting a first component of a specific binding pair with a second component of said pair, said first component being labeled with a molecule according to Formula (I) having a detectable group; and (b) monitoring said reaction by monitoring at least one of a fluorescence-emission intensity, a fluorescence lifetime, a fluorescence polarization, a fluorescence anisotropy, or fluorescence correlation of said detectable group.
- 57. An assay method for monitoring a binding process, comprising:
(a) reacting a first component of a specific binding pair with a second component of said pair, said first component being labeled with a molecule according to Formula (I) wherein X of Formula (I) is a fluorochrome, and said second component containing Z, wherein Z is one of a fluorochrome and a chromophore, Z being able to participatein fluorescence energy transfer, fluorescence quenching, or exciton formation with X; and (b) monitoring said reaction by monitoring a fluorescent property of said X.
- 58. The assay according to claim 57, wherein said fluorescent property is at least one of fluorescence-emission intensity, a fluorescence lifetime, a fluorescence polarization, a fluorescence anisotropy, or a fluorescence correlation.
- 59. An assay method for monitoring a binding process, comprising:
(a) reacting a first component of a specific binding pair with a second component of said pair, said first component being labeled with a molecule according to Formula (I) wherein X of Formula (I) is selected from the group consisting of a fluorochrome and a chromophore, and said second component containing Z, wherein Z is a fluorochrome able to participate in fluorescent energy transfer, fluorescence quenching, or exciton formation with said X; and (b) monitoring the reaction by monitoring a fluorescent property of said Z.
- 60. The assay according to claim 59, wherein said fluorescent property is at least one of fluorescence-emission intensity, a fluorescence lifetime, a fluorescence polarization, a fluorescence anisotropy, or a fluorescence correlation.
- 61. An assay method for monitoring a reaction, comprising:
(a) reacting a first participant in a reaction with a second participant in said reaction, said first participant being labeled with a molecule according to Formula (I); and (b) monitoring said reaction by monitoring a change in at least one detectable property of said detectable group.
- 62. The assay according to claim 61, wherein said detectable property is at least one of a fluorescent emission intensity, a fluorescence lifetime, a fluorescence polarization, a fluorescence anisotropy, and a fluorescence correlation.
- 63. A method for isolating at least one target material of interest comprising:
(a) contacting at least one molecule according to Formula (I) immobilized on a solid support, with a solution containing a target material having a target sequence of the form: C(X)iC(X)jC(X)kC, wherein C is cysteine, X is any amino acid, and i, j, and k are each independently 0 or an integer of from 1 to 6, under conditions that allow binding of said polypeptide to said immobilized molecule of Formula (I); and (b) eluting said target material with a low-molecular weight monothiol or low-molecular-weight dithiol.
- 64. The method according to claim 63, further comprising the step of washing said solid support to remove unbound material before eluting said target material.
- 65. The method according to claim 63, wherein said solid support is selected from the group consisting of a surface, a bead, a gel, and a chromatographic matrix.
- 66. A method for immobilizing at least one target material of interest including:
(a) contacting at least one molecule according to Formula (I) immobilized on a solid support with a solution containing a target material having a target sequence of the form: C(X)iC(X)jC(X)kC, wherein C is cysteine, X is any amino acid, and i, j, and k are each independently 0 or an integer of from 1 to 6, under conditions that allow binding of said target material to said immobilized molecule of Formula (I).
- 67. The method of claim 66, further comprising the step of washing said solid support to remove unbound material.
- 68. The method according to claim 66, wherein said solid support is selected from the group consisting of a surface, a bead, a gel, and a chromatographic matrix.
- 69. A method of screening a library of peptides and identifying a cysteine-containing peptide that binds to a molecule according to Formula (I) having a detectable group, the method comprising:
(a) incubating said molecule of Formula (I) with a peptide library, wherein said library comprises a multiplicity of solid supports, each solid support bearing a single species of peptide, under conditions that permit formation of a complex between said molecule according to Formula (I) and a peptide having a target sequence; and (b) detecting said detectable group, thereby identifying a solid support bearing a species of peptide that binds said molecule of Formula (I).
- 70. The method according to claim 69, wherein said method further comprises determining or inferring a sequence of the species of peptide on said solid support.
- 71. A method of screening a library of peptides and identifying a cysteine-containing peptide that binds to a molecule according to Formula (I) having a detectable group, the method comprising:
(a) introducing said molecule of Formula (I) to a peptide library, wherein said library comprises a multiplicity of peptides arrayed on a surface; and (b) identifying a location on the surface having a peptide thereon that binds the molecule of Formula (I) by detecting said detectable group.
- 72. The method according to claim 71, further comprising determining or inferring a sequence of a peptide at said location.
- 73. A method of screening a library of peptides and identifying a peptide that binds to a molecule according to Formula (I), the method comprising:
contacting molecules of Formula (I) immobilized on a solid support with a peptide library, wherein said library comprises a multiplicity of particles, each of said particles containing a single species of peptide, and also containing a nucleic acid that encodes at least one amino acid of said peptide.
- 74. The method according to claim 73, wherein said particles are nm-scale particles.
- 75. The method according to claim 73, wherein said particles are μm-scale particles.
- 76. The method according to claim 73, wherein said particles are selected from the group consisting of polysomes, bacteriophages, and cells.
- 77. The method according to claim 73, further comprising the step of eluting said material.
- 78. The method according to claim 73, further comprising the steps of: washing said solid support to remove unbound material; and eluting said material.
- 79. The method according to claim 77, wherein said eluting step includes using a low-molecular weight monothiol or a low-molecular weight dithiol.
- 80. The method according to claim 77, further comprising using said eluted material to repeat the procedure in claim 73.
- 81. The method according to claim 73, further comprising the step of determining or inferring an amino acid sequence of said target sequence containing peptide by determining a nucleotide sequence of said nucleic acid encoding at least one amino acid of said peptide.
- 82. A kit comprising:
(a) a molecule according to Formula (I); and (b) a molecule containing a target sequence, said target sequence comprising an amino acid sequence of the form: C(X)iC(X)jC(X)kC, wherein C is cysteine, X is any amino acid, and i, j, and k are each independently 0 or an integer of from 1 to 6.
- 83. A kit comprising:
(a) a molecule according to Formula (I); and (b) a reagent that promotes the formation of a complex between said molecule according to Formula (I) and a peptide comprising a target sequence, said target sequence comprising an amino acid sequence of the form C(X)iC(X)jC(X)kC, wherein C is cysteine, X is any amino acid, and i, j, and k are each independently 0 or an integer of from 1 to 6.
- 84. The method of synthesis of a compound of claim 1 by coupling:
(a) a synthon selected from the group consisting of mono-phenylarsine-functionalized 2,3,3-trimethylindole, mono-phenylarsine-functionalized 2,3,3-trimethylbenzindole, mono-phenylarsine-functionalized 2-methyl-pyridine, mono-phenylarsine-functionalized 2-methyl-benzothiazole, mono-phenylarsine-functionalized 2-methyl-napthothiazole, mono-phenylarsine-functionalized 2-methyl-benzoxazole, and mono-phenylarsine-functionalized 2-methyl-napthoxazole; (b) a synthon, identical or nonidentical to the synthon in (a), selected from the group in (a); and (c) a synthon containing at least one carbon atom.
- 85. The method of claim 84, wherein said coupling is performed as a single reaction step.
- 86. The method of claim 84, wherein said coupling comprises: either (i) first reacting (a) and (c) to form a product, followed by further reacting the product with (b); or (ii) first reacting (b) and (c) to form a product, followed by further reacting the product with (a).
- 87. The method of synthesis of a compound of claim 1 by coupling:
(a) a synthon selected from the group consisting of mono-phenylarsine-functionalized 2,3,3-trimethyl-5-sulfanato-indole, mono-phenylarsine-functionalized 2,3,3-trimethyl-6-sulfanato-benzindole, mono-phenylarsine-functionalized 2-methyl-5-sulfanato-pyridine, mono-phenylarsine-functionalized 2-methyl-5-sulfanato-benzothiazole, mono-phenylarsine-functionalized 2-methyl-6-sulfanato-napthothiazole, mono-phenylarsine-functionalized 2-methyl-5-sulfanato-benzoxazole, and mono-phenylarsine-functionalized 2-methyl-6-sulfanato-napthoxazole; (b) a synthon, identical or nonidentical to the synthon in (a), selected from the group in (a); and (c) a synthon containing at least one carbon atom.
- 88. The method of claim 87, wherein said coupling is performed as a single reaction step.
- 89. The method of claim 87, wherein said coupling comprises: either (i) first reacting (a) and (c) to form a product, followed by further reacting the product with (b); or (ii) first reacting (b) and (c) to form a product, followed by further reacting the product with (a).
- 90. The method of synthesis of a compound of claim 1 by coupling:
(a) a synthon selected from the group consisting of mono-phenylarsine-functionalized 2,3,3-trimethylindole, mono-phenylarsine-functionalized 2,3,3-trimethylbenzindole, mono-phenylarsine-functionalized 2-methyl-pyridine, mono-phenylarsine-functionalized 2-methyl-benzothiazole, mono-phenylarsine-functionalized 2-methyl-napthothiazole, mono-phenylarsine-functionalized 2-methyl-benzoxazole, and mono-phenylarsine-functionalized 2-methyl-napthoxazole; (b) a synthon selected from the group consisting of mono-phenylarsine-functionalized 2,3,3-trimethyl-5-sulfanato-indole, mono-phenylarsine-functionalized 2,3,3-trimethyl-6-sulfanato-benzindole, mono-phenylarsine-functionalized 2-methyl-5-sulfanato-pyridine, mono-phenylarsine-functionalized 2-methyl-6-sulfanato-benzothiazole, mono-phenylarsine-functionalized 2-methyl-6-sulfanato-napthothiazole, mono-phenylarsine-functionalized 2-methyl-5-sulfanato-benzoxazole, and mono-phenylarsine-functionalized 2-methyl-6-sulfanato-napthoxazole; and (c) a synthon containing at least one carbon atom.
- 91. The method of claim 90, wherein said coupling is performed as a single reaction step.
- 92. The method of claim 90, wherein said coupling comprises: either (i) first reacting (a) and (c) to form a product, followed by further reacting the product with (b); or (ii) first reacting (b) and (c) to form a product, followed by further reacting the product with (a).
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application claims priority to U.S. Provisional Patent Application No. 60/388,699, filed on Jun. 14, 2002, which is herein incorporated by reference in its entirety.
Government Interests
[0002] This invention was made with Government support under Grant No. NIH R01-GM41376, awarded by the National Institutes of Health. Therefore, the Government has certain rights in this invention.
Provisional Applications (1)
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Number |
Date |
Country |
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60388699 |
Jun 2002 |
US |