RT Journal Article
SR Electronic
T1 Developing NMR methods for macromolecular machines: Measurement of residual dipolar couplings to probe dynamic regions of the ribosome
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 319251
DO 10.1101/319251
A1 Xiaolin Wang
A1 John P. Kirkpatrick
A1 Hélène M. M. Launay
A1 Alfonso de Simone
A1 Daniel Häussinger
A1 Christopher M. Dobson
A1 Michele Vendruscolo
A1 Lisa D. Cabrita
A1 Christopher A. Waudby
A1 John Christodoulou
YR 2018
UL http://biorxiv.org/content/early/2018/07/12/319251.abstract
AB We describe an NMR approach based on the measurement of residual dipolar couplings (RDCs) to probe the structural and motional properties of the dynamic regions of the ribosome. Alignment of intact 70S ribosomes in filamentous bacteriophage enabled measurement of RDCs in the stalk protein bL12, and this information was used to refine a 3D structure of its C-terminal domain (CTD). Orientational constraints on the CTD alignment arising from the semiflexible linker sequence were further probed by a paramagnetic alignment strategy, and provided direct experimental validation of a structural ensemble previously derived from SAXS and NMR relaxation measurements. Our results demonstrate the prospect of better characterising dynamical and functional regions of more challenging macromolecular machines and systems, for example ribosome–nascent chain complexes.