Lipid probes and uses thereof
Disclosed herein are methods, compositions, probes, assays and kits for identifying a lipid binding protein as a drug binding target. Also disclosed herein are methods, compositions, and probes for mapping a ligand binding site on a lipid binding protein, identification of lipid binding proteins, generating drug-lipid binding protein profiles, high throughput drug screening, and identification of drugs as potential lipid binding protein ligands.
1. A process of generating a lipid probe-protein composition, comprising:
contacting a sample comprising a protein with a lipid probe; and
treating the sample and the lipid probe by a photoreactive means to generate the lipid probe-protein composition, wherein the treating time is from about 5 minutes to about 1 hour; and
wherein the lipid probe is a lipid probe of Formula (I):
wherein:
A is C 16 -C 20 alkyl or C 16 -C 20 alkenyl;
D is —NH 2 , —NHR 7 , or —OR 8 ;
R 7 is C 1 -C 4 alkyl, (C 1 -C 5 alkyl)OH, or (C 1 -C 5 )SO 3 M;
R 8 is (CH 2 OH)n;
M is monovalent or divalent cation;
n is 1, 2, or 3;
PG is a photoreactive group; and
AH is an affinity handle; and
wherein PG is attached to A or D and AH is attached to A or D.
2. The process of claim 1 , wherein the treating time is about 10 minutes.
3. The process of claim 1 , wherein the photoreactive group comprises azides, benzophenone, diazo compounds, diazirines, diazonium salts, or diaryl ketones.
4. The process of claim 1 , wherein the affinity handle comprises a carbodiimide, N-hydroxysuccinimide (NHS) ester, imidoester, pentafluorophenyl ester, hydroxymethyl phosphine, maleimide, haloacetyl, pyridyl disulfide, thiosulfonate, vinylsulfone, hydrazide, alkoxyamine, alkyne, azide, or isocyanate group.
5. The process of claim 1 , wherein the protein is an enzyme, a transporter, a receptor, an adaptor, a channel protein, or a chaperone.
6. The process of claim 1 , wherein the protein is encoded by a gene of Table 3, a protein encoded by a gene of Table 4, or a protein encoded by a gene of Table 5.
7. The process of claim 1 , wherein the photoreactive means comprises ultraviolet light.
8. The process of claim 1 , wherein the treating occurs at a temperature of from about 0° C. to about 40° C.
9. The process of claim 1 , wherein the treating occurs at a temperature of about 4° C.
10. A lipid probe-protein composition generated by the process of claim 2 .
11. A process of generating a lipid probe-protein composition, comprising:
contacting a sample comprising a protein with a lipid probe, wherein the lipid probe is selected from lipid probes 1, 3-10, 12-15, 17-21 and 23 of Table 1; and
treating the sample and the lipid probe by a photoreactive means to generate the lipid probe-protein composition, wherein the treating time is from about 5 minutes to about 1 hour.
12. The process of claim 11 , wherein the treating time is about 10 minutes.
13. The process of claim 11 , wherein the photoreactive group comprises azides, benzophenone, diazo compounds, diazirines, diazonium salts, or diaryl ketones.
14. The process of claim 11 , wherein the affinity handle comprises a carbodiimide, N-hydroxysuccinimide (NHS) ester, imidoester, pentafluorophenyl ester, hydroxymethyl phosphine, maleimide, haloacetyl, pyridyl disulfide, thiosulfonate, vinylsulfone, hydrazide, alkoxyamine, alkyne, azide, or isocyanate group.
15. The process of claim 11 , wherein the protein is an enzyme, a transporter, a receptor, an adaptor, a channel protein, or a chaperone.
16. The process of claim 11 , wherein the protein is encoded by a gene of Table 3, a protein encoded by a gene of Table 4, or a protein encoded by a gene of Table 5.
17. The process of claim 11 , wherein the photoreactive means comprises ultraviolet light.
18. The process of claim 11 , wherein the treating occurs at a temperature of from about 0° C. to about 40° C.
19. The process of claim 11 , wherein the treating occurs at a temperature of about 4° C.
20. A lipid probe-protein composition generated by the process of claim 11 .