title: Chemical development of intracellular protein heterodimerizers.
abstract: Cell activation initiated by receptor ligands or oncogenes triggers complex and convoluted intracellular signaling. Techniques initiating signals at defined starting points and cellular locations are attractive to elucidate the output of selected pathways. Here, we present the development and validation of a protein heterodimerization system based on small molecules cross-linking fusion proteins derived from HaloTags and SNAP-tags. Chemical dimerizers of HaloTag and SNAP-tag (HaXS) show excellent selectivity and have been optimized for intracellular reactivity. HaXS force protein-protein interactions and can translocate proteins to various cellular compartments. Due to the covalent nature of the HaloTag-HaXS-SNAP-tag complex, intracellular dimerization can be easily monitored. First applications include protein targeting to cytoskeleton, to the plasma membrane, to lysosomes, the initiation of the PI3K/mTOR pathway, and multiplexed protein complex formation in combination with the rapamycin dimerization system.
authors.affiliation: Department of Biomedicine, University of Basel, 4003 Basel, Switzerland.
authors.forename: Dominik
authors.initials: D
authors.lastname: Erhart
authors.forename: Mirjam
authors.initials: M
authors.lastname: Zimmermann
authors.forename: Olivier
authors.initials: O
authors.lastname: Jacques
authors.forename: Matthias B
authors.initials: MB
authors.lastname: Wittwer
authors.forename: Beat
authors.initials: B
authors.lastname: Ernst
authors.forename: Edwin
authors.initials: E
authors.lastname: Constable
authors.forename: Marketa
authors.initials: M
authors.lastname: Zvelebil
authors.forename: Florent
authors.initials: F
authors.lastname: Beaufils
authors.forename: Matthias P
authors.initials: MP
authors.lastname: Wymann
mesh_terms: D019883:Alkyl and Aryl Transferases / Q000378:metabolism; D000818:Animals; D002460:Cell Line; D003432:Cross-Linking Reagents / Q000737:chemistry / Q000378:metabolism*; D003599:Cytoskeleton / Q000378:metabolism; D019281:Dimerization; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D006867:Hydrolases / Q000378:metabolism; D051379:Mice; D041681:NIH 3T3 Cells; D019869:Phosphatidylinositol 3-Kinases / Q000737:chemistry / Q000378:metabolism; D054730:Protein Interaction Domains and Motifs; D011506:Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D015398:Signal Transduction; D013379:Substrate Specificity; D058570:TOR Serine-Threonine Kinases / Q000737:chemistry / Q000378:metabolism
chemical_list: D019883:Alkyl and Aryl Transferases; D003432:Cross-Linking Reagents; D049452:Green Fluorescent Proteins; D006867:Hydrolases; D019869:Phosphatidylinositol 3-Kinases; D011506:Proteins; D011994:Recombinant Proteins; D058570:TOR Serine-Threonine Kinases; C050956:DNA alkyltransferase; C046137:haloalkane dehalogenase