Measurement of Internal Movements within the 30S Ribosomal Subunit Using Förster Resonance Energy Transfer
- 25 November 2005
- journal article
- Published by Elsevier BV in Journal of Molecular Biology
- Vol. 354 (2), 459-472
- https://doi.org/10.1016/j.jmb.2005.09.010
Abstract
We have used Förster resonance energy transfer (FRET) to study specific conformational changes in the Escherichia coli 30 S ribosomal subunit that occur upon association with the 50 S subunit. By measuring energy transfer between 13 different pairs of fluorescent probes attached to specific positions on 30 S subunit proteins, we have monitored changes in distance between different locations within the 30 S subunit in its free and 50 S-bound states. The measured distance changes provide restraints for modeling the movement that occurs within the 30 S subunit upon formation of the 70 S ribosome in solution. Treating the head, body, and platform domains of the 30 S subunit as simple rigid bodies, the lowest-energy solution converges on a model that satisfies each of the individual FRET restraints. In this model, the 30 S subunit head tilts towards the 50 S subunit, similar to the movement found in comparing 30 S subunits and 70 S ribosomes from X-ray and cryo-electron microscope structures, and the platform is predicted to undergo a clock-wise rotation upon association.Keywords
Funding Information
- Ford Foundation
- W. M. Keck Foundation
- National Institutes of Health (PHS 5P41-RR03155)
- Damon Runyon Cancer Research Foundation
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