Molding atomic structures into intermediate-resolution cryo-EM density maps of ribosomal complexes using real-space...

Real-space refinement has been previously introduced as a flexible fitting method to interpret medium-resolution cryo-EM density maps in terms of atomic structures. In this way, conformational changes related to functional processes can be analyzed on the molecular level. In the application of the technique to the ribosome, quasiatomic models have been derived that have advanced our understanding of translocation. In this article, the choice of parameters for the fitting procedure is discussed. The quality of the fitting depends critically on the number of rigid pieces into which the model is divided. Suitable quality indicators are crosscorrelation, R factor, and density residual, all of which can also be locally applied. The example of the ribosome may provide some guidelines for general applications of real-space refinement to flexible fitting problems.