Serial X-ray liquidography: multi-dimensional assay framework for exploring biomolecular structural dynamics with microgram quantities

Seong Ok Kim, So Ri Yun, Hyosub Lee, Junbeom Jo, Doo-Sik Ahn, Doyeong Kim, Irina Kosheleva, Robert Henning, Jungmin Kim, Changin Kim, Seyoung You, Hanui Kim, Sang Jin Lee and Hyotcherl Ihee ()
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Seong Ok Kim: Institute for Basic Science (IBS)
So Ri Yun: Institute for Basic Science (IBS)
Hyosub Lee: Institute for Basic Science (IBS)
Junbeom Jo: Institute for Basic Science (IBS)
Doo-Sik Ahn: Institute for Basic Science (IBS)
Doyeong Kim: Institute for Basic Science (IBS)
Irina Kosheleva: The University of Chicago, 9700 South Cass Avenue
Robert Henning: The University of Chicago, 9700 South Cass Avenue
Jungmin Kim: Institute for Basic Science (IBS)
Changin Kim: Institute for Basic Science (IBS)
Seyoung You: Institute for Basic Science (IBS)
Hanui Kim: Institute for Basic Science (IBS)
Sang Jin Lee: Institute for Basic Science (IBS)
Hyotcherl Ihee: Institute for Basic Science (IBS)

Nature Communications, 2024, vol. 15, issue 1, 1-13

Abstract: Abstract Understanding protein structure and kinetics under physiological conditions is crucial for elucidating complex biological processes. While time-resolved (TR) techniques have advanced to track molecular actions, their practical application in biological reactions is often confined to reversible photoreactions within limited experimental parameters due to inefficient sample utilization and inflexibility of experimental setups. Here, we introduce serial X-ray liquidography (SXL), a technique that combines time-resolved X-ray liquidography with a fixed target of serially arranged microchambers. SXL breaks through the previously mentioned barriers, enabling microgram-scale TR studies of both irreversible and reversible reactions of even a non-photoactive protein. We demonstrate its versatility in studying a wide range of biological reactions, highlighting its potential as a flexible and multi-dimensional assay framework for kinetic and structural characterization. Leveraging X-ray free-electron lasers and micro-focused X-ray pulses promises further enhancements in both temporal resolution and minimizing sample quantity. SXL offers unprecedented insights into the structural and kinetic landscapes of molecular actions, paving the way for a deeper understanding of complex biological processes.

Date: 2024
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DOI: 10.1038/s41467-024-50696-0

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