Spin-spin coupling measurements in solid-state NMR and applications to rhodospin

Abstract

This thesis discusses some recently developed symmetry-based rotor-synchronised re-coupling sequences in magic-angle-spinning solid-state NMR and their applications to biological samples.

Symmetry theorems are applied to the problem of spin-spin couplings in the presence of magic-angle-spinning in the solid-state.  Homonuclear dipolar and J-coupling measurements are achieved on multiple ¹³C labelled systems.  J-coupling measurements have been performed on a G-protein coupled receptor - rhodopsin.  Accurate distance measurements and J-coupling are obtained through rare spin signal build-up curves and J-modulated curves.  The data collected for the G- protein coupled receptor includes the use of a new technique without decoupling.

A general introduction to solid-state NMR and its uses in molecular structure determination are discussed.  This includes a description of symmetry-based pulse sequences.  The emphasis is put on applications of such sequences for the establishment of an accurate methodology to measure spin-spin couplings.  The techniques are demonstrated on model systems of known structure and then applied to the investigation of the chemical bonding in the membrane protein rhodopsin, where the retinylidene chromophore is selectively labelled.

In the context of this thesis the main focus is on a particular class of biomolecule, the trans-membrane G-protein coupled receptor, rhodopsin.  A general description of the functionality of these proteins is also discussed.

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