Purification of ribosomes from human cell lines
- PMID: 21154551
- DOI: 10.1002/0471143030.cb0340s49
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.
Similar articles
-
A reconstituted cell-free assay for the evaluation of the intrinsic activity of purified human ribosomes.Nat Protoc. 2016 Jul;11(7):1309-25. doi: 10.1038/nprot.2016.072. Epub 2016 Jun 23. Nat Protoc. 2016. PMID: 27336708
-
Rapid purification of ribosomal particles assembled on histone H4 mRNA: a new method based on mRNA-DNA chimaeras.Biochem J. 2013 Feb 1;449(3):719-28. doi: 10.1042/BJ20121211. Biochem J. 2013. PMID: 23140180
-
Ribosome purification approaches for studying interactions of regulatory proteins and RNAs with the ribosome.Methods Mol Biol. 2012;905:273-89. doi: 10.1007/978-1-61779-949-5_18. Methods Mol Biol. 2012. PMID: 22736011 Free PMC article.
-
Guarding the 'translation apparatus': defective ribosome biogenesis and the p53 signaling pathway.Wiley Interdiscip Rev RNA. 2011 Jul-Aug;2(4):507-22. doi: 10.1002/wrna.73. Epub 2011 Jan 20. Wiley Interdiscip Rev RNA. 2011. PMID: 21957040 Review.
-
Ten remarks on peptide bond formation on the ribosome.Biochem Soc Trans. 2005 Jun;33(Pt 3):493-8. doi: 10.1042/BST0330493. Biochem Soc Trans. 2005. PMID: 15916550 Review.
Cited by
-
TGF-β2 Induces Ribosome Activity, Alters Ribosome Composition and Inhibits IRES-Mediated Translation in Chondrocytes.Int J Mol Sci. 2024 May 5;25(9):5031. doi: 10.3390/ijms25095031. Int J Mol Sci. 2024. PMID: 38732249 Free PMC article.
-
Diphthamide deficiency promotes association of eEF2 with p53 to induce p21 expression and neural crest defects.Nat Commun. 2024 Apr 26;15(1):3301. doi: 10.1038/s41467-024-47670-1. Nat Commun. 2024. PMID: 38671004 Free PMC article.
-
ERMA (TMEM94) is a P-type ATPase transporter for Mg2+ uptake in the endoplasmic reticulum.Mol Cell. 2024 Apr 4;84(7):1321-1337.e11. doi: 10.1016/j.molcel.2024.02.033. Epub 2024 Mar 20. Mol Cell. 2024. PMID: 38513662
-
Depletion of SNORA33 Abolishes ψ of 28S-U4966 and Affects the Ribosome Translational Apparatus.Int J Mol Sci. 2023 Aug 8;24(16):12578. doi: 10.3390/ijms241612578. Int J Mol Sci. 2023. PMID: 37628759 Free PMC article.
-
Customization of the translational complex regulates mRNA-specific translation to control CNS regeneration.Neuron. 2023 Sep 20;111(18):2881-2898.e12. doi: 10.1016/j.neuron.2023.06.005. Epub 2023 Jul 12. Neuron. 2023. PMID: 37442131 Free PMC article.
MeSH terms
LinkOut - more resources
Full Text Sources
