Application of nuclear magnetic resonance spectroscopy to the structure determination of the integral membrane proteins of the Mer operon

Efforts at elucidating the structural biology of membrane proteins represent an ongoing challenge to conventional methods of structure determination. The emergence of new methods for the measurement and application of orientational restraints have offered new avenues of pursuing the determination of membrane protein structures. Presented in this thesis is the evolution of experimental and computation methods necessary to extend NMR based structure determination methods to the polytopic mercuric ion transport proteins of the mer operon. Primary structural efforts are focused upon the bi-spanning protein, MerF, using solution-state NMR methods on protein reconstituted into isotropic and weakly aligned micelles and solid-state NMR methods on protein reconstituted into statically aligned bicelles. The application of the methods developed for MerF are applied to a tri-spanning chimeric protein, MerT f, to extend the NMR based methodology toward the structure determination of the principal mercuric ion transporter, MerT.