title: Chemical Genetics Approach to Engineer Kinesins with Sensitivity towards a Small-Molecule Inhibitor of Eg5.
abstract: Due to their fast and often reversible mode of action, small molecules are ideally suited to dissect biological processes. Yet, the validity of small-molecule studies is intimately tied to the specificity of the applied compounds, thus imposing a great challenge to screens for novel inhibitors. Here, we applied a chemical-genetics approach to render kinesin motor proteins sensitive to inhibition by the well-characterized small molecule S-Trityl-l-cysteine (STLC). STLC specifically inhibits the kinesin Eg5 through binding to a known allosteric site within the motor domain. Transfer of this allosteric binding site into the motor domain of the human kinesins Kif3A and Kif4A sensitizes them towards STLC. Single-molecule microscopy analyses confirmed that STLC inhibits the movement of chimeric but not wild-type Kif4A along microtubules. Thus, our proof-of-concept study revealed that this chemical-genetic approach provides a powerful strategy to specifically inhibit kinesins in vitro for which small-molecule inhibitors are not yet available.
authors.affiliation: Department of Biology, University of Konstanz, Universitätsstrasse 10, 78457, Konstanz, Germany.
authors.forename: Martin M
authors.identifier: 0000-0003-1237-2410
authors.initials: MM
authors.lastname: Möckel
authors.affiliation: Department of Biology, University of Konstanz, Universitätsstrasse 10, 78457, Konstanz, Germany.|FGen GmbH, Hochbergerstrasse 60C, 4057, Basel, Switzerland.
authors.forename: Corinna
authors.initials: C
authors.lastname: Hund
authors.affiliation: Department of Biology, University of Konstanz, Universitätsstrasse 10, 78457, Konstanz, Germany. thomas.u.mayer@uni-konstanz.de.
authors.forename: Thomas U
authors.identifier: 0000-0001-6954-2287
authors.initials: TU
authors.lastname: Mayer
mesh_terms: D055162:Biocatalysis; D004791:Enzyme Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D006801:Humans; D016547:Kinesins / Q000037:antagonists & inhibitors* / Q000737:chemistry / Q000378:metabolism; D008958:Models, Molecular; D015202:Protein Engineering; D054852:Small Molecule Libraries / Q000737:chemistry / Q000494:pharmacology*
chemical_list: D004791:Enzyme Inhibitors; C470243:KIF11 protein, human; D054852:Small Molecule Libraries; D016547:Kinesins
keywords: allosterism; chemical genetics; inhibitors; kinesin; mechanochemistry