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Chemical development of intracellular protein heterodimerizers
Journal
Chemistry & Biology
Volume
20
Number
4
Pages / Article-Number
549-57
Keywords
Alkyl and Aryl Transferases/metabolism; Animals; Cell Line; Cross-Linking Reagents/chemistry/ metabolism; Cytoskeleton/metabolism; Dimerization; Green Fluorescent Proteins/genetics/metabolism; HEK293 Cells; HeLa Cells; Humans; Hydrolases/metabolism; Mice; NIH 3T3 Cells; Phosphatidylinositol 3-Kinases/chemistry/metabolism; Protein Interaction Domains and Motifs; Proteins/chemistry/genetics/ metabolism; Recombinant Proteins/biosynthesis/chemistry/genetics; Signal Transduction; Substrate Specificity; TOR Serine-Threonine Kinases/chemistry/metabolism
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.
