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
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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。
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CiteScore
7.30
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