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

Abstract

AbstractThe 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.