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Dynamic post-translational histone modifications studied by NMR spectroscopy

通过核磁共振波谱研究动态翻译后组蛋白修饰

基本信息

批准号：
BB/H022570/1
负责人：
Flemming Hansen
金额：
$ 166.39万
依托单位：
University College London
依托单位国家：
英国
项目类别：
Fellowship
财政年份：
2010
资助国家：
英国
起止时间：
2010 至无数据
项目状态：
已结题

来源：
https://gtr.ukri.org/projects?ref=BB%2FH022570%2F1
关键词：
Dynamic post translational histone modifications

项目摘要

DNA molecules of human cells are many times longer than the diameter of the cell and consequently the DNA is packed into a compact structure called the chromatin. The chromatin consists of DNA molecules coiled around histone proteins (sticky pulleys) in a very systematic manner. The cell utilises several mechanisms to control exactly what inheritable information from the DNA molecule that is turned into functional product (cellular machines). One mechanism that the cell exploits is to change the charge of certain histone proteins (weaken or strengthen the stickiness of the pulleys) and thus expose or restrict a specific part of the DNA to the cells gene production apparatus. HDAC, an enzyme that is responsible for changing the charge of histone proteins (a stickiness enhancer) will be the focus of the proposed research project. The HDAC enzyme works as a scissors that strips a negative charge off the histone proteins, thereby rendering the histone tails positively charged which strengthen the interaction with the negatively charged DNA. In particular, the focus of the proposed project is the dynamics and molecular motions of the HDAC enzyme (how does the scissors cut) and the dynamics will be studied primarily with nuclear magnetic resonance (NMR) spectroscopy. Thus, one of the key objectives of the research is to characterise, at atomic resolution, the mechanism by which the HDAC enzyme alter the histone charges. The goal is also to characterise how HDAC enzymes interact with inhibitors (drugs) and histones. HDAC enzymes are involved in cancers where they are believed to suppress the production of tumour suppressors. Inhibitors of HDAC enzymes have shown anti-tumour activity and it is therefore likely that the outcome of the proposed research will lead to the design of specific inhibitors of HDAC enzymes ultimately resulting in more efficient cancer therapy. An understanding of histone modifications requires a detailed picture of the three-dimensional structure of the involved enzymes and an appreciation of how these structures vary and fluctuate with time (a scissors cuts due to its opening and closing motions). Static structures of HDAC enzymes have been determined over the last decade, however, very few studies on the flexibility and dynamics of these regulatory molecules have been published. The proposed project focuses on the use of NMR spectroscopy as the primary biophysical tool to elucidate molecular flexibility and interactions since NMR has the potential to provide a description of the dynamics and interactions at atomic resolution. It is the goal that the NMR measurements together with other experimental techniques and computer simulations will create a coherent characterisation of the enzyme function. Another major objective of the proposed research is to develop new NMR methods to characterise molecular dynamics and flexibility in general. These developments aim at a time-resolved description of enzyme motions, that is, a visualisation of the enzyme motions over time - as a movie - as opposed to previous methods that primarily provides the amplitudes of protein motions. The research will be carried out at the Institute of Structural and Molecular Biology (ISMB), a joint venture between Birkbeck and University College London (UCL). UCL and ISMB provide a state-of-the-art and stimulating research environment with dedicated NMR machines suitable for the proposed project. Also, the highly collaborative environment and world-class expertise at ISMB and UCL open up the possibility for fruitful collaborations. For example, to increase the likelihood of success in the development of new HDAC inhibitors, I have initiated a collaboration with Prof Charles Marson, UCL, who is an expert on the productions of HDAC inhibitors. In my opinion this collaboration will allow the results about the HDAC enzyme dynamics, obtained by NMR spectroscopy, to be taken one important step further towards the design of new medicine.

人类细胞的DNA分子比细胞的直径长许多倍，因此DNA被包装成称为染色质的紧凑结构。染色质由DNA分子以非常系统的方式盘绕在组蛋白（粘性滑轮）周围组成。细胞利用几种机制来精确地控制来自DNA分子的哪些可遗传信息转化为功能产物（细胞机器）。细胞利用的一种机制是改变某些组蛋白的电荷（减弱或加强滑轮的粘性），从而将DNA的特定部分暴露或限制在细胞的基因生产装置中。HDAC是一种负责改变组蛋白（一种粘性增强剂）电荷的酶，将成为拟议研究项目的重点。HDAC酶就像一把剪刀，可以剥离组蛋白上的负电荷，从而使组蛋白尾部带正电，从而加强与带负电荷的DNA的相互作用。特别是，拟议项目的重点是HDAC酶的动力学和分子运动（剪刀如何剪切），动力学将主要通过核磁共振（NMR）光谱进行研究。因此，该研究的关键目标之一是在原子分辨率下研究HDAC酶改变组蛋白电荷的机制。目标还包括表征HDAC酶如何与抑制剂（药物）和组蛋白相互作用。HDAC酶与癌症有关，据信它们抑制肿瘤抑制因子的产生。HDAC酶的抑制剂已显示出抗肿瘤活性，因此，拟议研究的结果可能会导致HDAC酶的特异性抑制剂的设计，最终导致更有效的癌症治疗。要理解组蛋白修饰，需要对所涉及的酶的三维结构有一个详细的了解，并了解这些结构如何随时间变化和波动（剪刀由于其开合运动而剪切）。在过去的十年中，HDAC酶的静态结构已经确定，然而，很少有关于这些调节分子的灵活性和动力学的研究已经发表。拟议项目的重点是使用核磁共振波谱作为主要的生物物理工具，以阐明分子的灵活性和相互作用，因为核磁共振有可能提供一个原子分辨率的动力学和相互作用的描述。目标是NMR测量与其他实验技术和计算机模拟一起将创建酶功能的一致表征。拟议研究的另一个主要目标是开发新的NMR方法来研究分子动力学和一般的灵活性。这些发展旨在对酶运动进行时间分辨的描述，即酶运动随时间的可视化-作为电影-与主要提供蛋白质运动幅度的先前方法相反。这项研究将在伯克贝克学院和伦敦大学学院（UCL）的合资企业结构和分子生物学研究所（ISMB）进行。UCL和ISMB提供了一个国家的最先进的和刺激的研究环境与专用的核磁共振机器适合拟议的项目。此外，ISMB和UCL的高度协作环境和世界一流的专业知识为富有成效的合作开辟了可能性。例如，为了增加新HDAC抑制剂开发成功的可能性，我与UCL的Charles Marson教授合作，他是HDAC抑制剂生产方面的专家。在我看来，这项合作将使通过NMR光谱学获得的HDAC酶动力学的结果朝着新药设计迈出重要的一步。

项目成果

期刊论文数量（10）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Protein NMR - Modern Techniques and Biomedical Applications

蛋白质 NMR - 现代技术和生物医学应用

DOI：
10.1007/978-1-4899-7621-5_3
发表时间：
2015
期刊：
影响因子：
0
作者：
Sauerwein A
通讯作者：
Sauerwein A

Solution structure of the major factor VIII binding region on von Willebrand factor.

冯·维勒布兰德因子上主要因子 VIII 结合区域的溶液结构。

DOI：
10.1182/blood-2013-07-517086
发表时间：
2014
期刊：
Blood
影响因子：
20.3
作者：
Shiltagh N
通讯作者：
Shiltagh N

Accurate structure and dynamics of the metal-site of paramagnetic metalloproteins from NMR parameters using natural bond orbitals.

DOI：
10.1021/ja209348p
发表时间：
2012-03-14
期刊：
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
影响因子：
15
作者：
Hansen, D. Flemming;Westler, William M.;Kunze, Micha B. A.;Markley, John L.;Weinhold, Frank;Led, Jens J.
通讯作者：
Led, Jens J.

Excited States of Nucleic Acids Probed by Proton Relaxation Dispersion NMR Spectroscopy.

DOI：
10.1002/anie.201605870
发表时间：
2016-09-19
期刊：
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
影响因子：
16.6
作者：
Juen, Michael Andreas;Wunderlich, Christoph Hermann;Nussbaumer, Felix;Tollinger, Martin;Kontaxis, Georg;Konrat, Robert;Hansen, D. Flemming;Kreutz, Christoph
通讯作者：
Kreutz, Christoph

Loop interactions and dynamics tune the enzymatic activity of the human histone deacetylase 8.