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.
Sabzian‐Molaei et al. (Wed,) studied this question.