Cryogen-Free Arbitrary Waveform EPR for Structural Biology and Biophysics

适用于结构生物学和生物物理学的无冷冻剂任意波形 EPR

基本信息

批准号：
BB/R013780/1
负责人：
Bela Bode
金额：
$ 26.63万
依托单位：
University of St Andrews
依托单位国家：
英国
项目类别：
Research Grant
财政年份：
2018
资助国家：
英国
起止时间：
2018 至无数据
项目状态：
已结题

来源：
https://gtr.ukri.org/projects?ref=BB%2FR013780%2F1
关键词：
Cryogen Free Arbitrary Waveform EPR

项目摘要

This equipment upgrade will enhance a large number of molecular biosciences related projects that utilise electron paramagnetic resonance (EPR) spectroscopy and will have a major impact in the biomedical sciences and structural biology at the Universities of St Andrews and Dundee and beyond.EPR is an ideal method for obtaining specific information at the nanoscale, as it is exquisitely sensitive to the magnetic spins of radicals and other paramagnetic centres. These are of interest as many are hotspots of biochemical activity. Recently, the number of applications utilising EPR has greatly increased. This increase was catalysed by new technology which allows researchers to selectively introduce spins into biomolecules and use them as molecular beacons. The magnetic moments of the spins interact with the magnetic moments of other paramagnets or magnetic nuclei and thus magnetically illuminate their surroundings. It has become increasingly popular to use measurements of the long-range distances between the spins to map the nanoworlds of protein conformations and interactions quite as if navigating by the relative positions and brightness of lighthouses.Recently, developments have started to transform this science. While spins are commonly manipulated by on/off electromagnetic irradiation, the advent of arbitrary waveform generators has opened up a whole new universe allowing access to new realms of information in experiments. This technology is often proposed to be the future of EPR and the analogous advancements have wholly transformed the field of nuclear magnetic resonance. Now that the more demanding technical requirements of EPR are fulfilled it seems a logical imperative to upgrade to this technology as the upgrade even just means a moderate additional cost compared to the initial investment.A second development concerns the need to cool the spins to extremely low temperatures using liquid helium. Helium is an expensive, finite resource that has proven to be highly susceptible to disruption in the supply chain and poses a considerable safety hazard. Traditionally, liquid helium would be connected to the spectrometer before performing EPR experiments. Recently, cryogen-free cryostats have been developed using closed-cycle cooling that operates like an ultra-low temperature refrigerator. In these cryostats, used helium is cooled by electrically driven compression and expansion cycles to re-liquefy and thus be recycled. This greatly improves reliability and sustainability. Significantly, this also reduces risks to the researchers whilst performing experiments. Additionally, the increased stability means that the facility can operate around the clock and for a longer period of time, allowing more experiments to be performed and more scientific questions to be answered in a given time, thereby substantially improving the efficiency of the facility.The Universities of St Andrews and Dundee have developed an extensive programme of biological applications using this methodology and now seek to implement cryogen-free arbitrary waveform EPR to enhance existing facilities for projects investigating biomedical challenges and opportunities in the bio-based economy of the future.

此次设备升级将增强大量利用电子顺磁共振（EPR）光谱的分子生物科学相关项目，并将对圣安德鲁斯大学和邓迪大学及其他大学的生物医学科学和结构生物学产生重大影响。EPR是一个理想的选择这是一种在纳米尺度上获取特定信息的方法，因为它对自由基和其他顺磁中心的磁自旋极其敏感。这些都很有趣，因为许多都是生化活动的热点。最近，利用 EPR 的申请数量大幅增加。这种增长是由新技术催化的，该技术允许研究人员选择性地将自旋引入生物分子并将其用作分子信标。自旋的磁矩与其他顺磁体或磁核的磁矩相互作用，从而磁性地照亮它们的周围环境。使用自旋之间的远距离测量来绘制蛋白质构象和相互作用的纳米世界已经变得越来越流行，就像通过灯塔的相对位置和亮度进行导航一样。最近，进展已经开始改变这门科学。虽然自旋通常是通过开/关电磁辐射来操纵的，但任意波形发生器的出现开辟了一个全新的宇宙，允许在实验中访问新的信息领域。这项技术经常被认为是 EPR 的未来，类似的进步已经彻底改变了核磁共振领域。既然 EPR 更苛刻的技术要求已经得到满足，那么升级到该技术似乎是合乎逻辑的必然，因为与初始投资相比，升级甚至只意味着适度的额外成本。第二个发展涉及将旋转冷却到极低的需要使用液氦的温度。氦气是一种昂贵且有限的资源，事实证明，它极易受到供应链中断的影响，并造成相当大的安全隐患。传统上，在进行 EPR 实验之前，会将液氦连接到光谱仪。最近，已经开发出使用闭式循环冷却的无制冷剂低温恒温器，其运行方式类似于超低温冰箱。在这些低温恒温器中，用过的氦气通过电力驱动的压缩和膨胀循环进行冷却，以重新液化，从而被回收利用。这极大地提高了可靠性和可持续性。值得注意的是，这也降低了研究人员在进行实验时的风险。此外，稳定性的提高意味着该设施可以全天候运行并运行更长的时间，从而可以在给定时间内进行更多的实验并回答更多的科学问题，从而大幅提高设施的效率。圣安德鲁斯大学和邓迪大学利用这种方法开发了广泛的生物应用计划，现在寻求实施无冷冻剂任意波形 EPR，以增强现有设施，用于研究未来生物经济中生物医学挑战和机遇的项目。

项目成果

期刊论文数量（9）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Investigating Native Metal Ion Binding Sites in Mammalian Histidine-Rich Glycoprotein

研究哺乳动物富含组氨酸的糖蛋白中的天然金属离子结合位点

DOI：
10.26434/chemrxiv-2023-f6n6p
发表时间：
2023
期刊：
影响因子：
0
作者：
Ackermann K
通讯作者：
Ackermann K

Investigating Native Metal Ion Binding Sites in Mammalian Histidine-Rich Glycoprotein.

DOI：
10.1021/jacs.3c00587
发表时间：
2023-04-12
期刊：
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
影响因子：
15
作者：
Ackermann, Katrin;Khazaipoul, Siavash;Wort, Joshua L.;Sobczak, Amelie I. S.;El Mkami, Hassane;Stewart, Alan J.;Bode, Bela E.
通讯作者：
Bode, Bela E.

Pulse dipolar EPR for determining nanomolar binding affinities.