title: Hypothemycin, a fungal natural product, identifies therapeutic targets in Trypanosoma brucei [corrected].
abstract: Protein kinases are potentially attractive therapeutic targets for neglected parasitic diseases, including African trypanosomiasis caused by the protozoan, Trypanosoma brucei. How to prioritize T. brucei kinases and quantify their intracellular engagement by small-molecule inhibitors remain unsolved problems. Here, we combine chemoproteomics and RNA interference to interrogate trypanosome kinases bearing a Cys-Asp-Xaa-Gly motif (CDXG kinases). We discovered that hypothemycin, a fungal polyketide previously shown to covalently inactivate a subset of human CDXG kinases, kills T. brucei in culture and in infected mice. Quantitative chemoproteomic analysis with a hypothemycin-based probe revealed the relative sensitivity of endogenous CDXG kinases, including TbGSK3short and a previously uncharacterized kinase, TbCLK1. RNAi-mediated knockdown demonstrated that both kinases are essential, but only TbCLK1 is fully engaged by cytotoxic concentrations of hypothemycin in intact cells. Our study identifies TbCLK1 as a therapeutic target for African trypanosomiasis and establishes a new chemoproteomic tool for interrogating CDXG kinases in their native context. DOI:http://dx.doi.org/10.7554/eLife.00712.001.
authors.affiliation: Tetrad Graduate Program , University of California, San Francisco , San Francisco , United States ; Center for Discovery and Innovation in Parasitic Diseases , University of California, San Francisco , San Francisco , United States.
authors.forename: Mari
authors.initials: M
authors.lastname: Nishino
authors.forename: Jonathan W
authors.initials: JW
authors.lastname: Choy
authors.forename: Nathan N
authors.initials: NN
authors.lastname: Gushwa
authors.forename: Juan A
authors.initials: JA
authors.lastname: Oses-Prieto
authors.forename: Kyriacos
authors.initials: K
authors.lastname: Koupparis
authors.forename: Alma L
authors.initials: AL
authors.lastname: Burlingame
authors.forename: Adam R
authors.initials: AR
authors.lastname: Renslo
authors.forename: James H
authors.initials: JH
authors.lastname: McKerrow
authors.forename: Jack
authors.initials: J
authors.lastname: Taunton
mesh_terms: D000345:Affinity Labels; D000981:Antiprotozoal Agents / Q000494:pharmacology*; D005658:Fungi / Q000378:metabolism*; D013058:Mass Spectrometry; D014346:Trypanosoma brucei brucei / Q000187:drug effects*; D015025:Zearalenone / Q000031:analogs & derivatives* / Q000494:pharmacology
chemical_list: D000345:Affinity Labels; D000981:Antiprotozoal Agents; C120440:hypothemycin; D015025:Zearalenone
keywords: Mouse; Other; T. brucei; chemoproteomics; hypothemycin; protein kinase