Photocleavable chemical inducers of dimerization (CID) and methods of use
The instant invention provides photolysable compounds, and their use in reversible chemical induced dimerization and light-induced regulation of proteins.
1. A compound represented by the formula R 1 —Y—R 2 , wherein
R 1 is a ligand capable of selectively binding to a first receptor;
R 2 is a ligand capable of selectively binding to a second receptor;
Y is a linker providing a covalent linkage between R 1 and R 2 ,
wherein Y is photocleavable;
R 1 is methotrexate, dexamethasone, 3,5,3′-triiodothyronine, trans-retinoic acid, biotin, coumermycin, tetracycline, lactose, FK506,
analogs or derivatives thereof;
R 2 is
analogs or derivatives thereof,
wherein X═O or NH;
Y is
2. The compound of claim 1 , wherein R 1 is a ligand capable of selectively binding to dihydrofolate reductase (DHFR).
3. The compound of claim 2 , wherein the ligand capable of selectively binding to dihydrofolate reductase is trimethoprim (TMP) or a derivative or analog thereof, or methotrexate or a derivative or analog thereof.
4. The compound of claim 2 , wherein R 2 is a ligand capable of selectively binding to FK506 Binding Protein (FKBP).
5. The compound of claim 4 , wherein the ligand capable of selectively binding to FK506 Binding Protein (FKBP) is a synthetic ligand for FKBP (SLF) or a derivative or analog thereof.
6. The compound of claim 1 , wherein Y is cleavable by exposure to light at a wavelength of about 405 nm.
7. The compound of claim 6 , wherein Y comprises a dimethoxynitrobenzyl (DMNB) moiety or a derivative or analog thereof.
8. A complex comprising a compound of claim 1 complexed to (i) a first protein domain comprising a first binding domain and (ii) a second protein domain comprising a second binding domain.
9. A method of regulating activity of a protein, the method comprising:
(a) providing a complex comprising
(i) a first protein domain comprising a first binding domain;
(ii) a second protein domain comprising a second binding domain; and
(iii) a compound of claim 1 ,
wherein the compound of claim 1 is bound to the first protein domain and the second protein domain;
(b) cleaving the compound of claim 1 by exposure to light, thereby cleaving the complex and regulating activity of the protein.
10. The method of claim 9 , wherein the first binding domain is dihydrofolate reductase (DHFR).
11. The method of claim 10 , wherein the DHFR is E. coli DHFR.
12. The method of claim 9 , wherein the second binding domain is FK506 binding protein (FKBP).
13. The method of claim 9 , wherein the first protein domain comprises one domain of an active protein.
14. The method of claim 9 , wherein the second protein domain comprises another domain of an active protein.
15. The method of claim 9 , wherein the first protein domain and the second protein domain together comprise a functional protein.
16. The method of claim 9 , wherein regulating the activity of a protein comprises inactivating the protein.
17. The method of claim 9 , wherein regulating the activity of a protein comprises activating the protein.
18. A method of regulating the activity of a protein in a cell, the method comprising:
(a) providing a cell that expresses a first protein domain comprising a first binding domain and a second protein domain that comprises a second binding domain;
(b) providing the compound of claim 1 that dimerizes the first and second protein domains; and
(c) cleaving the compound of claim 1 by exposure to light, thereby cleaving the complex and regulating activity of the protein.
19. The method of claim 18 , wherein dimerization of the first and second protein domains produces an active protein.
20. The method of claim 18 , wherein dimerization of the first and second protein domains inactivates the protein.