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Chromatographic Purification of Highly Active Yeast Ribosomes
Published on: October 24, 2011
A novel chromatography system to isolate active ribosomes from pathogenic bacteria.
Bruce A Maguire1, Lillian M Wondrack, Leonard G Contillo
1Department of Exploratory Medicinal Sciences, Pfizer Global Research and Development, Groton Laboratories, Groton, Connecticut 06340, USA. Bruce.Maguire@pfizer.com
RNA (New York, N.Y.)
|November 14, 2007
Summary
A new chromatography method rapidly isolates active bacterial ribosomes. This technique preserves ribosome integrity, enabling better functional assays for developing novel antibiotics targeting bacterial ribosomes.
Area of Science:
- Microbiology
- Biochemistry
- Drug Discovery
Background:
- Ribosomes are crucial targets for antibiotic development.
- Current methods for ribosome isolation are often harsh, damaging ribosome structure and function.
- Intact, active ribosomes are needed for accurate functional assays.
Purpose of the Study:
- To develop a novel, rapid chromatography method for isolating active bacterial ribosomes.
- To preserve ribosome integrity during isolation.
- To enable improved functional assays for antibiotic discovery.
Main Methods:
- Development of a novel chromatography technique utilizing an alkyl linker attached to a resin.
- Interaction of ribosomes with the resin, primarily through their RNA component.
- Application of the method to various bacterial species, including clinical isolates.
Main Results:
- Rapid isolation of active ribosomes without harsh conditions.
- Demonstrated success with Escherichia coli, Deinococcus radiodurans, Streptococcus pneumoniae, and MRSA.
- Obtained unusually intact and highly active ribosomes.
Conclusions:
- The novel chromatography method provides intact, active ribosomes from diverse bacterial sources.
- This advancement significantly improves in vitro functional assays.
- Facilitates the discovery and development of new ribosome-targeted antibiotics.
