Advancing time-resolved structural biology: latest strategies in cryo-EM and X-ray crystallography.

Banari, Amir; Samanta, Amit K; Munke, Anna; Laugks, Tim; Bajt, Saša; Grünewald, Kay; Marlovits, Thomas C; Küpper, Jochen et al. · Nat Methods · 2025

review · Level V

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Abstract

Structural biology offers a window into the functionality of molecular machines in health and disease. A fundamental challenge lies in capturing both the high-resolution structural details and dynamic changes that are essential for function. The high-resolution methods of X-ray crystallography and electron cryo-microscopy (cryo-EM) are mainly used to study ensembles of static conformations but can also capture crucial dynamic transition states. Here, we review the latest strategies and advancements in time-resolved structural biology with a focus on capturing dynamic changes. We describe recent technology developments for time-resolved sample preparation and delivery in the cryo-EM and X-ray fields and explore how these technologies could mutually benefit each other to advance our understanding of biology at the molecular and atomic scales.

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