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A light-triggered time-resolved x-ray solution scattering (tr-xss) workflow with application to protein conformational dynamics
Umeå University, Faculty of Science and Technology, Department of Chemistry.
Umeå University, Faculty of Science and Technology, Department of Chemistry.ORCID iD: 0000-0002-0706-7414
Umeå University, Faculty of Science and Technology, Department of Chemistry.
Umeå University, Faculty of Science and Technology, Department of Chemistry.ORCID iD: 0000-0002-3364-6647
2026 (English)In: FEBS Open Bio, E-ISSN 2211-5463Article in journal (Refereed) Epub ahead of print
Abstract [en]

Time-resolved X-ray solution scattering (TR-XSS) is a synchrotron-based methodology that enables real-time structural characterization under near-native conditions to provide insight into dynamic and transient structural changes inaccessible to static high-resolution methods such as cryo-electron microscopy (cryo-EM) or X-ray crystallography. Here, we present a workflow for light-triggered TR-XSS experiments that spans data collection, data processing, kinetic analysis, and structural refinement, with accompanying Python scripts. A calcium-transporting P-type ATPase membrane protein (LMCA1) is used as an illustrative example, but the protocol is broadly applicable to diverse protein systems. This workflow offers a practical framework for collecting TR-XSS synchrotron data and subsequent data analysis and interpretation.

Place, publisher, year, edition, pages
John Wiley & Sons, 2026.
Keywords [en]
kinetic modeling, protein conformational dynamics, structural refinement, time-resolved X-ray solution scattering (TR-XSS)
National Category
Biochemistry Molecular Biology
Identifiers
URN: urn:nbn:se:umu:diva-250064DOI: 10.1002/2211-5463.70200PubMedID: 41669764Scopus ID: 2-s2.0-105029833005OAI: oai:DiVA.org:umu-250064DiVA, id: diva2:2041832
Funder
EU, Horizon 2020, 101081419Swedish Research Council, 2024-04385The Kempe Foundations, JCSMK 24-543Available from: 2026-02-26 Created: 2026-02-26 Last updated: 2026-02-26

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Sabzian-Molaei, FatemehOrädd, FredrikMagkakis, KonstantinosAndersson, Magnus

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