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.
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Acknowledgments
The original protocol of the 80S ribosome purification and crystal treatments was elaborated by Adam Ben-Shem. The data collection strategy was suggested by Clemens Schulze-Briese and Marcus Mueller, beam scientists at the SLS, x06sa. The work on the 80S structure determination has been greatly assisted by the authors of original publications and was supported by awards from EMBO, HFSP, CNRS, ANR and SPINE2.
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Melnikov, S. (2013). Structure of the Eukaryotic Ribosome: Tips and Tricks. In: Read, R., Urzhumtsev, A., Lunin, V. (eds) Advancing Methods for Biomolecular Crystallography. NATO Science for Peace and Security Series A: Chemistry and Biology. Springer, Dordrecht. https://doi.org/10.1007/978-94-007-6232-9_27
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DOI: https://doi.org/10.1007/978-94-007-6232-9_27
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