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A Bioorthogonal Small‐Molecule‐Switch System for Controlling Protein Function in Live Cells |
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| Authors: | Dr Peng Liu Abram Calderon Dr Georgios Konstantinidis Dr Jian Hou Dipl Stephanie Voss Dr Xi Chen Fu Li Soumya Banerjee Dr Jan‐Erik Hoffmann Christiane Theiss Dr Leif Dehmelt Dr Yao‐Wen Wu |
| Institution: | 1. Chemical Genomics Centre of the Max Planck Society, Otto‐Hahn‐Str. 15, 44227 Dortmund (Germany) http://www.cgc.mpg.de/index.php/research‐groups/rg‐dr‐yaowen‐wu/research;2. Abteilung Physikalische Biochemie, Max‐Planck‐Institut für molekulare Physiologie, Otto‐Hahn‐Str. 11, 44227, Dortmund (Germany);3. Abteilung Systemische Zellbiologie, Max‐Planck‐Institut für molekulare Physiologie (Germany);4. Chemische Biologie, Technische Universit?t Dortmund, Otto‐Hahn‐Str. 6, 44227 Dortmund (Germany) |
| Abstract: | Chemically induced dimerization (CID) has proven to be a powerful tool for modulating protein interactions. However, the traditional dimerizer rapamycin has limitations in certain in vivo applications because of its slow reversibility and its affinity for endogenous proteins. Described herein is a bioorthogonal system for rapidly reversible CID. A novel dimerizer with synthetic ligand of FKBP′ (SLF′) linked to trimethoprim (TMP). The SLF′ moiety binds to the F36V mutant of FK506‐binding protein (FKBP) and the TMP moiety binds to E. coli dihydrofolate reductase (eDHFR). SLF′‐TMP‐induced heterodimerization of FKBP(F36V) and eDHFR with a dissociation constant of 0.12 μM . Addition of TMP alone was sufficient to rapidly disrupt this heterodimerization. Two examples are presented to demonstrate that this system is an invaluable tool, which can be widely used to rapidly and reversibly control protein function in vivo. |
| Keywords: | cell protrusion dimerization intracellular translocation fluorescence proteins |