Trimethoprim-Halotag (TMP-HTag) is a small molecule chemical linker developed for the rapid and reversible control of protein localization in living cells (Ballister). TMP is an dihydrofolate reductase (DHFR) inhibitor chosen for its specificity in binding to the bacterial form of DHFR.[1][2] The other half of the linker is a Halotag, a self labelling bacterial globular protein ligand that can bind covalently and irreversibly to the chloroalkane group of a Haloenzyme.[2][3] Positioned between the TMP group and HaloTag is a flexible linker that can be modified to optimize protein linking efficiency.[2][3] The modular structure of TMP-HaloTag makes it an ideal heterobifunctional tool for use in chemically induced dimerization (CID).[1][2] Additionally, TMP- HTag can be modified to include photo-cleavable groups that allow for the control of CID using light.[1][2]
NVOC group between TMP and HTag for photo cleavability. Requires light activation and use may result in photo toxicity. Light sensitive so must be stored and applied in controlled light. Slightly faster synthesis process than other dimerizer syntheses
Additional aromatic rings for rigidity that improves permeability and stability of heterodimer protein complexes that form with dimerization. Not light sensitive but slower. Lengthier synthesis but simpler process than TNH
Aromatic fluorines lend lipophilicity and possibly “stickiness” that facilitates faster dimerization than with TBH. Minimizes photo-toxicity since light is not required.
