title: Cell-permeant and photocleavable chemical inducer of dimerization.
abstract: Chemical inducers of dimerization (CIDs) have been developed to orchestrate protein dimerization and translocation. Here we present a novel photocleavable HaloTag- and SNAP-tag-reactive CID (MeNV-HaXS) with excellent selectivity and intracellular reactivity. Excitation at 360 nm cleaves the methyl-6-nitroveratryl core of MeNV-HaXS. MeNV-HaXS covalently links HaloTag- and SNAP-tag fusion proteins, and enables targeting of selected membranes and intracellular organelles. MeNV-HaXS-mediated translocation has been validated for plasma membrane, late endosomes, lysosomes, Golgi, mitochondria, and the actin cytoskeleton. Photocleavage of MeNV-HaXS liberates target proteins and provides access to optical manipulation of protein relocation with high spatiotemporal and subcellular precision. MeNV-HaXS supports kinetic studies of protein dynamics and the manipulation of subcellular enzyme activities, which is exemplified for Golgi-targeted cargo and the assessment of nuclear import kinetics.
authors.affiliation: University of Basel, Department of Biomedicine, Mattenstrasse 28, Basel (Switzerland).
authors.forename: Mirjam
authors.initials: M
authors.lastname: Zimmermann
authors.forename: Ruben
authors.initials: R
authors.lastname: Cal
authors.forename: Elia
authors.initials: E
authors.lastname: Janett
authors.forename: Viktor
authors.initials: V
authors.lastname: Hoffmann
authors.forename: Christian G
authors.initials: CG
authors.lastname: Bochet
authors.forename: Edwin
authors.initials: E
authors.lastname: Constable
authors.forename: Florent
authors.initials: F
authors.lastname: Beaufils
authors.forename: Matthias P
authors.initials: MP
authors.lastname: Wymann
mesh_terms: D002462:Cell Membrane / Q000378:metabolism; D002463:Cell Membrane Permeability; D003600:Cytosol / Q000378:metabolism; D004705:Endocytosis / Q000502:physiology / Q000528:radiation effects; D006056:Golgi Apparatus / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism*; D006147:Guanine / Q000031:analogs & derivatives / Q000737:chemistry / Q000494:pharmacology; D006367:HeLa Cells; D006801:Humans; D007700:Kinetics; D008027:Light; D008247:Lysosomes / Q000378:metabolism; D008928:Mitochondria / Q000378:metabolism; D017319:Photosensitizing Agents / Q000737:chemistry / Q000494:pharmacology*; D055503:Protein Multimerization / Q000187:drug effects*; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: C485182:Cyan Fluorescent Protein; D017319:Photosensitizing Agents; D011993:Recombinant Fusion Proteins; C064976:O(6)-benzylguanine; D049452:Green Fluorescent Proteins; D006147:Guanine
keywords: chemical inducers of dimerization; fusion proteins; photocleavable linkers; photolysis; protein-protein interactions