Structural studies of macromolecules in solution using small angle X-ray scattering

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dc.contributor.authorMrozowich, Tyler
dc.contributor.authorMcLennan, Steffane
dc.contributor.authorOverduin, Michael
dc.contributor.authorPater, Trushar R.
dc.date.accessioned2024-06-19T22:33:00Z
dc.date.available2024-06-19T22:33:00Z
dc.date.issued2018
dc.descriptionOpen access article. Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported license (CC BY-NC-NC 3.0) applies
dc.description.abstractProtein-protein interactions involving proteins with multiple globular domains present technical challenges for determining how such complexes form and how the domains are oriented/positioned. Here, a protocol with the potential for elucidating which specific domains mediate interactions in multicomponent system through ab initio modeling is described. A method for calculating solution structures of macromolecules and their assemblies is provided that involves integrating data from small angle X-ray scattering (SAXS), chromatography, and atomic resolution structures together in a hybrid approach. A specific example is that of the complex of full-length nidogen-1, which assembles extracellular matrix proteins and forms an extended, curved nanostructure. One of its globular domains attached to laminin y-1, which structures the basement membrane. This provides a basis for determining accurate structures of flexible multidomain protein complexes and is enabled by synchrotron sources coupled with automation robotics and size exclusion chromatography systems. This combination allows rapid analysis in which multiple oligomeric states are separated prior to SAXS data collection. The analysis yields information on the radius of gyration, particle dimension, molecular shape and interdomain pairing. The protocol for generating 3D models of complexes by fitting high-resolution structures of the component proteins is also given.
dc.identifier.citationMrozowich, T., McLennan, S., Overduin, M., & Patel, T. R. (2018). Structural studies of macromolecules in solution using small angle X-ray scattering. Journal of Visual Experiments, (141), Article e58538. https://www.jove.com/video/58538
dc.identifier.urihttps://hdl.handle.net/10133/6791
dc.language.isoen
dc.publisher.departmentDepartment of Chemistry and Biochemistry
dc.publisher.facultyArts and Science
dc.publisher.institutionUniversity of Lethbridge
dc.publisher.institutionUniversity of Alberta
dc.publisher.institutionUniversity of Calgary
dc.subjectBiochemistry
dc.subjectExtracellular matrix protein
dc.subjectHybrid solution structure
dc.subjectIaminin
dc.subjectMultidomain protein
dc.subjectNidogen
dc.subjectProtein interaction
dc.subjectProtein structure
dc.subjectStructural domain
dc.subjectSmall angle X-ray scattering
dc.subjectThree-dimension model
dc.subject.lcshExtracellular matrix proteins
dc.subject.lcshProtein-protein interactions
dc.subject.lcshThree-dimensional modeling
dc.titleStructural studies of macromolecules in solution using small angle X-ray scattering
dc.typeArticle
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