Affinity purification of ribosomes to access the translatome.

RE Halbeisen; T Scherrer; André P. Gerber

doi:10.1016/j.ymeth.2009.04.003

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Journal article

Affinity purification of ribosomes to access the translatome.

RE Halbeisen, T Scherrer and André P. Gerber

Methods, Vol.48(3), pp.306-310

07/2009

DOI: https://doi.org/10.1016/j.ymeth.2009.04.003

Abstract

Cell Fractionation

Affinity

Protein Array Analysis

RNA-Binding Proteins

Ribosomal Proteins

Ribosomes

Saccharomyces cerevisiae

Saccharomyces cerevisiae Proteins

Chromatography

We describe ribosome affinity purification (RAP), a method that allows rapid purification of ribosomes and associated messages from the yeast Saccharomyces cerevisiae. The method relies on the expression of protein A tagged versions of the ribosomal protein Rpl16, which is used to efficiently recover endogenously formed ribosomes and polysomes from cellular extracts with IgG-coupled spherical microbeads. This approach can be applied to profile reactions of the translatome, which refers to all messages associated with ribosomes, with those of the transcriptome using DNA microarrays. In addition, ribosomal proteins, their modifications, and/or other associated proteins can be mapped with mass spectrometry. Finally, application of this method in other organisms provides a valuable tool to decipher cell-type specific gene expression patterns.