High-resolution cryo-EM of the human CDK-activating kinase for structure-based drug design.
Victoria I CushingAdrian Fujiet KohJunjie FengKaste JurgaityteAlexander BondkeSebastian H B KrollMarion BarbazangesBodo ScheiperAsh K BahlAnthony G M BarrettSimak AliAbhay KotechaBasil J GreberPublished in: Nature communications (2024)
Rational design of next-generation therapeutics can be facilitated by high-resolution structures of drug targets bound to small-molecule inhibitors. However, application of structure-based methods to macromolecules refractory to crystallization has been hampered by the often-limiting resolution and throughput of cryogenic electron microscopy (cryo-EM). Here, we use high-resolution cryo-EM to determine structures of the CDK-activating kinase, a master regulator of cell growth and division, in its free and nucleotide-bound states and in complex with 15 inhibitors at up to 1.8 Å resolution. Our structures provide detailed insight into inhibitor interactions and networks of water molecules in the active site of cyclin-dependent kinase 7 and provide insights into the mechanisms contributing to inhibitor selectivity, thereby providing the basis for rational design of next-generation therapeutics. These results establish a methodological framework for the use of high-resolution cryo-EM in structure-based drug design.
Keyphrases
- high resolution
- small molecule
- electron microscopy
- cell cycle
- mass spectrometry
- protein kinase
- signaling pathway
- endothelial cells
- tyrosine kinase
- high speed
- tandem mass spectrometry
- adverse drug
- drug induced
- single molecule
- emergency department
- transcription factor
- pluripotent stem cells
- electronic health record
- liquid chromatography