Ribosomes and cryo-EM: a duet.

Area of Science:

• Structural biology
• Molecular biology
• Biophysics

Background:

• Ribosomes are essential molecular machines for protein synthesis.
• X-ray crystallography was the primary method for high-resolution ribosome structure determination.
• Electron cryomicroscopy (cryo-EM) has recently advanced to rival and exceed X-ray crystallography for ribosome studies.

Purpose of the Study:

• To highlight the advantages of cryo-EM over X-ray crystallography for ribosome structural analysis.
• To describe novel cryo-EM sample preparation and data processing techniques.
• To showcase new biological insights gained from cryo-EM ribosome structures.

Main Methods:

• Advancements in direct-electron detectors for cryo-EM.
• Innovative cryo-EM sample preparation strategies.
• Sophisticated cryo-EM data processing pipelines.

Main Results:

• Cryo-EM now achieves resolutions comparable to or better than X-ray crystallography for ribosomes.
• New cryo-EM approaches enable structural analyses not feasible with X-ray crystallography.
• Cryo-EM structures have revealed novel ribosome functions and mechanisms, including new polypeptide synthesis pathways and roles in supercomplexes.

Conclusions:

• Cryo-EM has become the leading technique for atomic-resolution ribosome structure determination.
• Cryo-EM offers unique analytical capabilities that expand our understanding of ribosome biology.
• Recent cryo-EM studies provide unprecedented insights into ribosome function across diverse biological contexts.