Large Temperature-Jump and Nanosecond Hyperquenching for Time-Resolved Structural Studies

IF 6.1 Q1 CHEMISTRY, MULTIDISCIPLINARY
Dr. Alexey V. Cherepanov, Prof. Dr. Harald Schwalbe
{"title":"Large Temperature-Jump and Nanosecond Hyperquenching for Time-Resolved Structural Studies","authors":"Dr. Alexey V. Cherepanov,&nbsp;Prof. Dr. Harald Schwalbe","doi":"10.1002/cmtd.202200050","DOIUrl":null,"url":null,"abstract":"<p>The quest for atomic structures of microsecond reaction intermediates is at the frontline of modern biochemistry. Currently, there is a clear lack of experimental methods for preparing necessary time-resolved samples. Here, we report the development of a single-turnover technique for nanosecond initiation and suspension of biomolecular reactions with kinetic resolution in the microsecond time domain. Reactions can be started by large temperature-jump or direct mixing and arrested by hyperquenching in liquid cryogen at a target temperature of 77 K. Diverse morphology of nanoscale glassy bodies feature among others thin field-of-view plane sheets that can be used for structure analyses of freeze-trapped macromolecules by transmission electron cryomicroscopy. We also report the ultra-high vacuum sublimation at 77 K – a novel method for concentrating reaction intermediates for structural studies by low-temperature techniques.</p>","PeriodicalId":72562,"journal":{"name":"Chemistry methods : new approaches to solving problems in chemistry","volume":"3 3","pages":""},"PeriodicalIF":6.1000,"publicationDate":"2023-01-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cmtd.202200050","citationCount":"1","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Chemistry methods : new approaches to solving problems in chemistry","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/cmtd.202200050","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"CHEMISTRY, MULTIDISCIPLINARY","Score":null,"Total":0}
引用次数: 1

Abstract

The quest for atomic structures of microsecond reaction intermediates is at the frontline of modern biochemistry. Currently, there is a clear lack of experimental methods for preparing necessary time-resolved samples. Here, we report the development of a single-turnover technique for nanosecond initiation and suspension of biomolecular reactions with kinetic resolution in the microsecond time domain. Reactions can be started by large temperature-jump or direct mixing and arrested by hyperquenching in liquid cryogen at a target temperature of 77 K. Diverse morphology of nanoscale glassy bodies feature among others thin field-of-view plane sheets that can be used for structure analyses of freeze-trapped macromolecules by transmission electron cryomicroscopy. We also report the ultra-high vacuum sublimation at 77 K – a novel method for concentrating reaction intermediates for structural studies by low-temperature techniques.

Abstract Image

时间分辨结构研究中的大温度跃变和纳秒超淬火
对微秒反应中间体的原子结构的探索处于现代生物化学的前沿。目前,明显缺乏制备必要的时间分辨样品的实验方法。在这里,我们报告了在微秒时间域具有动力学分辨率的生物分子反应的纳秒启动和悬浮的单次翻转技术的发展。反应可以通过大的温度跳跃或直接混合开始,并通过在液冷剂中以77 K的目标温度进行超淬火来阻止。纳米级玻璃体的不同形态特征之一是薄视场平面片,可用于通过透射电子冷冻显微镜对冻结大分子进行结构分析。我们还报道了77 K的超高真空升华——一种利用低温技术浓缩反应中间体用于结构研究的新方法。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 求助全文
来源期刊
CiteScore
7.30
自引率
0.00%
发文量
0
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信