A general approach to unraveling structural changes of food proteins using atomistic model-based SAXS

Small-angle X-ray scattering (SAXS) is widely used to probe macromolecular structures in solution, but conventional analysis often relies on low-resolution models that overlook chemical and molecular details essential for understanding protein behavior in biological and food systems. We present a methodology that integrates high-resolution SAXS modeling with AlphaFold-predicted oligomeric assemblies to enable hypothesis-driven structural interpretation under complex and physiologically relevant conditions. Using pea globulin as a model system, we test an hybrid approach across a temperature gradient to capture transitions from native globular assemblies to unfolded conformations. Association is described using structure factors, and thermally unfolded states are represented through rigid-body refinements. This strategy may provide more precise characterization of proteins across diverse conformational conditions. Furthermore, by applying a structure factor it could offer deeper insight into oligomerization, unfolding, and processing-related phase transitions. By bridging computational prediction with experimental scattering data, this approach allows for testing at high resolution a variety of structural assumptions, and could improve our understanding of the structural changes occurring due to environmental stimuli.