title: Purification of ribosomes from human cell lines.
abstract: Highly conserved during evolution, the ribosome is the central effector of protein synthesis. In mammalian cells, the ribosome is a macromolecular complex composed of four different ribosomal RNAs (rRNA) and about 80 ribosomal proteins. Requiring more than 200 factors, ribosome biogenesis is a highly complex process that takes place mainly within the nucleoli of eukaryotic cells. Crystallographic data suggest that the ribosome is a ribozyme, in which the rRNA catalyses the peptide bond formation and ensures quality control of the translation. Ribosomal proteins are involved in this molecular mechanism; nonetheless, their role is still not fully characterized. Recent studies suggest that ribosomes themselves and/or the mechanisms underlying their synthesis, processing, and assembly play a key role in the establishment and progression of several human pathologies. The protocol described here is simple, efficient, and robust, and allows one to purify high-quality ribosomes from human cultured cell lines. Ribosomes purified with this protocol are adequate for most of the subsequent analyses of their RNA and protein content.
authors.affiliation: Université de Lyon, Lyon, France.
authors.forename: Stéphane
authors.initials: S
authors.lastname: Belin
authors.forename: Sabine
authors.initials: S
authors.lastname: Hacot
authors.forename: Lionel
authors.initials: L
authors.lastname: Daudignon
authors.forename: Gabriel
authors.initials: G
authors.lastname: Therizols
authors.forename: Stéphane
authors.initials: S
authors.lastname: Pourpe
authors.forename: Hichem C
authors.initials: HC
authors.lastname: Mertani
authors.forename: Manuel
authors.initials: M
authors.lastname: Rosa-Calatrava
authors.forename: Jean-Jacques
authors.initials: JJ
authors.lastname: Diaz
mesh_terms: D002458:Cell Fractionation / Q000379:methods*; D002460:Cell Line; D006801:Humans; D014176:Protein Biosynthesis; D012270:Ribosomes / Q000235:genetics / Q000378:metabolism*