Interpretation of medium resolution cryoEM maps of multi-protein complexes

Highlights • CryoEM maps at medium (3.5–6 Å) resolution can be challenging to interpret. • Integration of multiple methods can inform cryoEM studies. • Mass spectrometry and biochemistry facilitate map interpretation and model building.
Electron cryo-microscopy (cryoEM) is used to determine structures of biological molecules, including multi-protein complexes. Maps at better than 3.0 Å resolution are relatively straightforward to interpret since atomic models of proteins and nucleic acids can be built directly. Still, these resolutions are often difficult to achieve, and map quality frequently varies within a structure. This results in data that are challenging to interpret, especially when crystal structures or suitable homology models are not available. Recent advances in mass spectrometry techniques, computational methods and model building tools facilitate subunit/domain fitting into maps, elucidation of protein contacts, and de novo generation of atomic models. Here, we review techniques for map interpretation and provide examples from recent studies of multi-protein complexes.