Engineered kinesin motor proteins amenable to small-molecule inhibition

• Published in: Nature communications Vol. 7; no. 1; p. 11159
• Main Authors: Engelke, Martin F, Winding, Michael, Yue, Yang, Shastry, Shankar, Teloni, Federico, Reddy, Sanjay, Blasius, T Lynne, Soppina, Pushpanjali, Hancock, William O, Gelfand, Vladimir I, Verhey, Kristen J
• Format: Journal Article
• Language: English
• Published: England Nature Publishing Group 05.04.2016; Nature Portfolio
• Subjects: Animals; Cell Line; Cell Movement - drug effects; Chlorocebus aethiops; COS Cells; Drosophila melanogaster; Dyneins - genetics; Dyneins - metabolism; Gene Expression; Humans; Kinesins - antagonists & inhibitors; Kinesins - genetics; Kinesins - metabolism; Mice; Microtubules - drug effects; Microtubules - metabolism; Microtubules - ultrastructure; Molecular Imaging; Myosins - genetics; Myosins - metabolism; Plasmids - chemistry; Plasmids - metabolism; Protein Engineering; Protein Transport; Recombinant Proteins - genetics; Recombinant Proteins - metabolism; Small Molecule Libraries - chemical synthesis; Small Molecule Libraries - pharmacology; Transfection; Tubulin Modulators - chemical synthesis; Tubulin Modulators - pharmacology
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/ncomms11159

The human genome encodes 45 kinesin motor proteins that drive cell division, cell motility, intracellular trafficking and ciliary function. Determining the cellular function of each kinesin would benefit from specific small-molecule inhibitors. However, screens have yielded only a few specific inhibitors. Here we present a novel chemical-genetic approach to engineer kinesin motors that can carry out the function of the wild-type motor yet can also be efficiently inhibited by small, cell-permeable molecules. Using kinesin-1 as a prototype, we develop two independent strategies to generate inhibitable motors, and characterize the resulting inhibition in single-molecule assays and in cells. We further apply these two strategies to create analogously inhibitable kinesin-3 motors. These inhibitable motors will be of great utility to study the functions of specific kinesins in a dynamic manner in cells and animals. Furthermore, these strategies can be used to generate inhibitable versions of any motor protein of interest.