title: T-CrAsH: a heterologous chemical crosslinker.
abstract: Copper-free click chemistry is currently the most promising and most rapidly developing technology for performing tailored chemical reactions inside intact living cells and animals. Its potential is particularly intensely explored in the field of live cell imaging, for both proteins and metabolites. Here we expand the application spectrum of click reactions to the chemical crosslinking of two proteins of choice in living cells. By combining strain-promoted Diels-Alder cycloaddition with FlAsH-based labeling of peptidic tetracysteine motifs, we developed the membrane-permeating reversible crosslinker T-CrAsH. We demonstrate the feasibility of the method both in vitro and inside cells. The biggest advantage of this new tool is the small size of the crosslinkable groups; this significantly decreases the risk of functional interference.
authors.affiliation: Cell Biology and Biophysics Unit, EMBL, Meyerhofstrasse 1, 69117 Heidelberg (Germany).
authors.forename: Anna
authors.initials: A
authors.lastname: Rutkowska
authors.forename: Tilman
authors.initials: T
authors.lastname: Plass
authors.forename: Jan-Erik
authors.initials: JE
authors.lastname: Hoffmann
authors.forename: Dmytro A
authors.initials: DA
authors.lastname: Yushchenko
authors.forename: Suihan
authors.initials: S
authors.lastname: Feng
authors.forename: Carsten
authors.initials: C
authors.lastname: Schultz
mesh_terms: D000596:Amino Acids / Q000737:chemistry; D057930:Click Chemistry; D003432:Cross-Linking Reagents / Q000138:chemical synthesis / Q000737:chemistry*; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D015394:Molecular Structure; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism
chemical_list: D000596:Amino Acids; D003432:Cross-Linking Reagents; D011506:Proteins
keywords: biarsenicals; click chemistry; crosslinkers; cycloaddition; protein interactions