A new frontier exists in unraveling interactive biological and biochemical processes and pathways at the macromolecular level. Of critical importance is the three-dimensional visualization of membrane proteins and molecular machines that control the trafficking trough cell compartment in their native functional state. Three techniques play a major role, NMR, XRD and Cryo-TEM.
Nuclear magnetic resonance (NMR) has the capability to study specific protein domains or fragments and their functional role in protein folding and dynamics and in ligand binding whereas X-Ray crystallography (XRD) allows visualizing high-resolution but more static 3D structures of apo and liganded proteins, mainly in a monomeric or dimeric state after crystallization. To unravel more physiologically relevant situations however, it is essential to visualize multimeric complexes in their tertiary and quaternary state and their interaction with other complexes. Cryo-TEM applications like single particle analysis one can visualize multimeric complexes. In this so-called translational methodology, cryo-TEM thus provides complementary information to NMR and XRD that can be crucial for a detailed structural analysis for a better understanding of the mechanism of the physiologically relevant complex.
Latest developments in the cryo-TEM workflow have brought the 3 major structural biology technologies closer together. Now, finally, a continuum has been reached on all important aspects with regards to resolution and macromolecular scales which allows for the full deployment of the combination of these technologies.
We will discuss the future of structural biology based on the latest developments of the Thermo Fisher workflow and its components.