Photocleavable linker
There are provided, inter alia, photolabile compounds and methods useful for the formation of dimers of biological molecules and subsequent dissociation of the dimers.
1. A method for forming a FK506 binding protein (FKBP) dimer, wherein each FKBP of the dimer is fused to a peptide, said method comprising:
(i) contacting a first FKBP fused to a peptide and a second FKBP fused to a peptide with a compound of Formula (I); and
(ii) allowing said compound to bind to said first FKBP fused to a peptide and said second FKBP fused to a peptide, thereby forming said FKBP dimer comprising a link between said first FKBP fused to a peptide and said second FKBP fused to a peptide;
thereby forming a FKBP dimer, wherein each FKBP of the dimer is fused to a peptide;
wherein,
said compound of Formula (I) has the structure
wherein,
L 1 is a photocleavable linker, and
z 1 and z 2 are each independently an integer in the range 0 to 6.
2. The method of claim 1 , wherein L 1 is substituted or unsubstituted alkylene, substituted or unsubstituted heteroalkylene, substituted or unsubstituted cycloalkylene, substituted or unsubstituted heterocycloalkylene, substituted or unsubstituted arylene or substituted or unsubstituted heteroarylene.
3. The method of claim 1 , wherein L 1 comprises a nitrophenylene.
4. The method of claim 1 , wherein said peptide is selected from the group consisting of a dynein intermediate chain (IC) peptide, a fragment of IC peptide, an analog of IC peptide, a dynein intermediate chain IC2C peptide, a fragment of dynein intermediate chain IC2C peptide, and an analog of dynein intermediate chain IC2C peptide.
5. The method of claim 4 , wherein said peptide is an IC peptide, or fragment or analog thereof.
6. The method of claim 5 , wherein said peptide binds LC8.
7. The method of claim 4 , wherein said peptide is an dynein intermediate chain IC2C peptide, or fragment or analog thereof.
8. The method of claim 7 , wherein said peptide binds TcTex1.
9. The method of claim 1 , wherein said first FKBP fused to a peptide and said second FKBP fused to a peptide further comprise a reporter group.
10. The method of claim 9 , wherein said reporter group is a dye, a radioactive moiety, or a protein.
11. A method for antagonizing a dynein-associated process in a cell, said method comprising:
(i) contacting a cell with the FK506 binding protein (FKBP) dimer of claim 1 under conditions suitable to antagonize a dynein-associated process.
12. The method of claim 11 , further comprising:
(ii) monitoring a dynein-associated process in said cell.
13. The method of claim 12 , wherein said monitoring monitors endosome dispersion, lysosome dispersion or Golgi dispersion.
14. The method of claim 13 , wherein said endosome dispersion is increased, lysosome dispersion is increased, or Golgi dispersion is increased.
15. The method of claim 13 , wherein endosome dispersion is monitored by assaying the level of early endosome markers 1 (EEA1).
16. The method of claim 13 , wherein Golgi dispersion is monitored by assaying the level of GM130 marker.
17. The method of claim 12 , further comprising
(iii) contacting said cell with light under conditions suitable to afford photocleavage of said compound of claim 1 .
18. The method of claim 12 , wherein said dynein-associated process is a cytoplasmic dynein function distinct from vesicular transport.
19. The method of claim 18 , said cytoplasmic dynein function is mitotic behavior.
20. A method for synthesizing a compound of Formula (IV):
said method comprising contacting a compound with structure of Formula (5)
with diol 6,
under conditions suitable to afford the compound of Formula (IV).