A novel chromatography system to isolate active ribosomes from pathogenic bacteria
Abstract
We have developed a novel chromatography for the rapid isolation of active ribosomes from bacteria without the use of harsh conditions or lengthy procedures that damage ribosomes. Ribosomes interact with an alkyl linker attached to the resin, apparently through their RNA component. Examples are given with ribosomes from Escherichia coli, Deinococcus radiodurans, and with clinical isolates of Streptococcus pneumoniae and methicillin-resistant Staphylococcus aureus (MRSA). The ribosomes obtained by this method are unusually intact, so that highly active ribosomes can now be isolated from the clinical isolates, enabling significantly improved in vitro functional assays that will greatly assist the discovery and development of new ribosomally targeted antibiotics.
Keywords
Footnotes
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Reprint requests to: Bruce A. Maguire, Department of Exploratory Medicinal Sciences, Pfizer Global Research and Development, Groton Laboratories, Groton, CT 06340, USA; e-mail: Bruce.Maguire{at}pfizer.com; fax: (860) 441-6090; or Zuoyu Xu, Bacteriology and Mycology Branch, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 6610 Rockledge Drive, Bethesda, MD 20892-6603, USA; e-mail: xuzuoyu{at}niaid.nih.gov; fax: (301) 402-2508.
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Abbreviations: rRNA, ribosomal RNA; TNT, transcription and translation; PAGE, polyacrylamide gel electrophoresis; IC50, inhibitor concentration that gives 50% inhibition; CPM, counts per minute; HIC, hydrophobic interaction chromatography; SEC, size exclusion chromatography; LC-MS, liquid chromatography mass spectrometry; CV, column volumes.
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Article published online ahead of print. Article and publication date are at http://www.rnajournal.org/cgi/doi/10.1261/rna.692408.
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- Received June 15, 2007.
- Accepted September 13, 2007.
- Copyright © 2008 RNA Society
