Open AccessIsolation of Bacterial Ribosomes with Monolith Chromatography
Author(s) -
Andrej Trauner,
Mark H. Bennett,
Huw D. Williams
Publication year - 2011
Publication title -
plos one
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.99
H-Index - 332
ISSN - 1932-6203
DOI - 10.1371/journal.pone.0016273
Subject(s) - ribosome , chromatography , ultracentrifuge , analytical ultracentrifugation , monolith , elution , isolation (microbiology) , sucrose gradient , ribosomal rna , ribosomal protein , chemistry , ion chromatography , biochemistry , biology , rna , microbiology and biotechnology , enzyme , gene , catalysis
We report the development of a rapid chromatographic method for the isolation of bacterial ribosomes from crude cell lysates in less than ten minutes. Our separation is based on the use of strong anion exchange monolithic columns. Using a simple stepwise elution program we were able to purify ribosomes whose composition is comparable to those isolated by sucrose gradient ultracentrifugation, as confirmed by quantitative proteomic analysis (iTRAQ). The speed and simplicity of this approach could accelerate the study of many different aspects of ribosomal biology.
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