CowScape: Quantitative reconstruction of the conformational landscape of biological macromolecules from cryo-EM data

원문 초록

Cryo-EM data processing typically focuses on the structure of the main conformational state under investigation and discards images that belong to other states. This approach can reach atomic resolution, but ignores vast amounts of valuable information about the underlying conformational ensemble and its dynamics. CowScape analyzes an entire cryo-EM dataset and thereby obtains a quantitative description of structural variability of macromolecular complexes that represents the biochemically relevant conformational space. By combining extensive image classification with principal component analysis (PCA) of the classified 3D volumes and kernel density estimation, CowScape can be used as a quantitative tool to analyze this variability. PCA projects all 3D structures along the major modes spanning a low-dimensional space that captures a large portion of structural variability. The number of particle images in a given state can be used to calculate an energy landscape based on kernel density estimation and Boltzmann inversion. By revealing allosteric interactions in macromolecular complexes, CowScape allows us to distinguish and interpret dynamic changes in macromolecular complexes during function and regulation.