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The Application of REDOR NMR to Understand the Conformation of Epothilone B
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...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5535963/ https://www.ncbi.nlm.nih.gov/pubmed/28698492 http://dx.doi.org/10.3390/ijms18071472 |
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author | Lee, Jae-Ho Kim, Moon-Su Lee, Hyo Won Lee, Ihl-Young C. Kim, Hyun Kyoung Kim, Nam Doo Lee, SangGap Seo, Hwajeong Paik, Younkee |
author_facet | Lee, Jae-Ho Kim, Moon-Su Lee, Hyo Won Lee, Ihl-Young C. Kim, Hyun Kyoung Kim, Nam Doo Lee, SangGap Seo, Hwajeong Paik, Younkee |
author_sort | Lee, Jae-Ho |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-5535963 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-55359632017-08-04 The Application of REDOR NMR to Understand the Conformation of Epothilone B Lee, Jae-Ho Kim, Moon-Su Lee, Hyo Won Lee, Ihl-Young C. Kim, Hyun Kyoung Kim, Nam Doo Lee, SangGap Seo, Hwajeong Paik, Younkee Int J Mol Sci Article 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. MDPI 2017-07-09 /pmc/articles/PMC5535963/ /pubmed/28698492 http://dx.doi.org/10.3390/ijms18071472 Text en © 2017 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Lee, Jae-Ho Kim, Moon-Su Lee, Hyo Won Lee, Ihl-Young C. Kim, Hyun Kyoung Kim, Nam Doo Lee, SangGap Seo, Hwajeong Paik, Younkee The Application of REDOR NMR to Understand the Conformation of Epothilone B |
title | The Application of REDOR NMR to Understand the Conformation of Epothilone B |
title_full | The Application of REDOR NMR to Understand the Conformation of Epothilone B |
title_fullStr | The Application of REDOR NMR to Understand the Conformation of Epothilone B |
title_full_unstemmed | The Application of REDOR NMR to Understand the Conformation of Epothilone B |
title_short | The Application of REDOR NMR to Understand the Conformation of Epothilone B |
title_sort | application of redor nmr to understand the conformation of epothilone b |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5535963/ https://www.ncbi.nlm.nih.gov/pubmed/28698492 http://dx.doi.org/10.3390/ijms18071472 |
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