Structure of the eukaryotic ribosome: Tips and tricks

Abstract

Ribosomes are biological assemblies consisting of more than 50 different proteins and several thousands of RNA bases. Recently the structure of the eukaryotic 80S ribosome was determined by methods of X-ray crystallography. This chapter highlights several steps and ideas of the experimental approach that can be applied for a broader range of biological complexes. The biochemistry for the 80S ribosome project (purification and crystal treatments) was derived from procedures and principles developed for structural studies of labile plant complexes and resulted in elaboration of a rapid and gentle purification protocol. The structure determination of this giant enzyme implied model building of 3.3MDa of RNA and protein moieties: remodeling of ∼2.3MDa residues of the conserved core, known from the structures of prokaryotic ribosomes, and de novo building of ∼1MDa of eukaryote-specific components. © 2013 Springer Science+Business Media Dordrecht.