1. The Application of REDOR NMR to Understand the Conformation of Epothilone B
- Author
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Jae-Ho Lee, Moon-Su Kim, Hyo Won Lee, Ihl-Young C. Lee, Hyun Kyoung Kim, Nam Doo Kim, SangGap Lee, Hwajeong Seo, and Younkee Paik
- Subjects
microtubules ,epothilone B ,REDOR ,solid-state NMR ,bioactive conformation ,Biology (General) ,QH301-705.5 ,Chemistry ,QD1-999 - Abstract
The structural information of small therapeutic compounds complexed in biological matrices is important for drug developments. However, structural studies on ligands bound to such a large and dynamic system as microtubules are still challenging. This article reports an application of the solid-state NMR technique to investigating the bioactive conformation of epothilone B, a microtubule stabilizing agent, whose analog ixabepilone was approved by the U.S. Food and Drug Administration (FDA) as an anticancer drug. First, an analog of epothilone B was designed and successfully synthesized with deuterium and fluorine labels while keeping the high potency of the drug; Second, a lyophilization protocol was developed to enhance the low sensitivity of solid-state NMR; Third, molecular dynamics information of microtubule-bound epothilone B was revealed by high-resolution NMR spectra in comparison to the non-bound epothilone B; Last, information for the macrolide conformation of microtubule-bound epothilone B was obtained from rotational-echo double-resonance (REDOR) NMR data, suggesting the X-ray crystal structure of the ligand in the P450epoK complex as a possible candidate for the conformation. Our results are important as the first demonstration of using REDOR for studying epothilones.
- Published
- 2017
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