From Slow to Ultra-fast MAS: Structural Determination of Type-Three Secretion System Bacterial Needles and Inorganic Materials by Solid-State NMR

by Jean-Philippe Demers
Doctoral thesis
Date of Examination:2014-04-23
Date of issue:2014-12-16
Advisor:Dr. Adam Lange
Referee:Prof. Dr. Konrad Samwer
Referee:Prof. Dr. Christian Griesinger
Referee:Prof. Dr. Helmut Grubmüller
Referee:Prof. Dr. Hans Christian Hofsäss
Referee:Prof. Dr. Markus Zweckstetter
Persistent Address: http://dx.doi.org/10.53846/goediss-4656

 

 

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Abstract

English

This work on solid-state NMR methodologies and applications is divided in three parts. In the first part, three methods for solid-state NMR at ultra-fast Magic-Angle Spinning (MAS) are introduced: completely low-power protein assignment experiments, broadband low-power cross-polarization, sensitivity enhancement by magnetization recovery. The second part present studies of inorganic materials where 11 compounds containing silicon, phosphorus and tin atoms are characterized. A new method employing ultra-fast Magi-Angle Spinning for the study of compounds with extremely large Chemical Shift Anisotropy (CSA) is introduced. The third part consists in the study of the needle of bacterial Type-Three Secretion Systems (T3SS). Following the study of the T3SS needle of Shigella flexneri, a common architecture for T3SS needles is identified. A novel hybrid structure determination approach combining solid-state NMR and cryo-electron microscopy was employed to produce the atomic-resolution structure of the T3SS needle of Shigella flexneri and resolve a controversy in the field of Type-Three secretion.
Keywords: Solid-state NMR; NMR pulse sequences; Inorganic chemistry; Type-Three Secretion System; Protein structure; Ultra-fast Magic-Angle Spinning; Shigella flexneri; Cryo-Electron Microscopy; Paramagnetic doping; Cross-polarization; Low-power radio-frequency pulses
 
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