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Type of Document Dissertation Author Evans, Michael Stanley URN etd-11302006-155501 Title CHAIN LENGTH-DEPENDENT CONFORMATIONS OF CO-TRANSLATIONAL FOLDING INTERMEDIATES OF P22 TAILSPIKE Degree Doctor of Philosophy Department Chemistry and Biochemistry Advisory Committee
Advisor Name Title Patricia L. Clark Committee Chair Anthony S. Serianni Committee Member Holly V. Goodson Committee Member Paul W. Huber Committee Member Keywords
- protein folding in vivo
- co-translational protein folding
- prokaryotic translation
Date of Defense 2006-11-27 Availability restricted Abstract Newly synthesized proteins must form their native structure in the crowded environment of the cell, while avoiding non-native conformations that can lead to aggregation. P22 tailspike is a homotrimer prone to aggregation via misfolding of its central beta-helix domain in vitro. To assess whether co-translational folding enables newly synthesized tailspike chains to avoid aggregation-prone conformations in vivo, a novel method was first developed to produce stalled ribosome nascent chain complexes. This new method was used to measure anti-tailspike monoclonal antibody binding to and partial protease digestion of four different tailspike nascent chain lengths. These experiments reveal ribosome-bound nascent tailspike chains populate ordered conformations with some native-state structural features, but these conformations are distinct from the predominant conformations of tailspike in vitro refolding intermediates and refolded, unstalled tailspike truncations. These results suggest the aggregation-prone beta-helix domain pre-organizes co-translationally, prior to chain release, and that this conformation is distinct from the global energy minimum for the truncated free chain in solution.Files
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