New methodologies in nuclear magnetic resonance (NMR) for the study of biological systems

In its relatively short history, Nuclear Magnetic Resonance (NMR) has become one of the most powerful analytical techniques available to the modern chemist, physicist, or biologist. Whether NMR methods are being applied to determine protein structures or probe the extent of spongiform degeneration in a patient with Creutzfeld-Jakob Disease (CJD), the field of NMR is constantly seeking to answer questions of vital biological and sociological interest. Through a greater understanding of protein structure, new treatments can be developed for diseases such as Alzheimer's Disease and what is considered to be the human variant of Mad Cow Disease. In this dissertation, several novel methods are presented to aid in structure determination of peptides and proteins.;Some of the new methods have focused upon structural elucidation methods in the solid state. Although the goal of these methods involved structure determination of the prion protein, the techniques presented in this dissertation could be employed on a variety of biological samples in the solid state. Further developments on a method for determining backbone (&phis;, y ) dihedral angles using 13Calpha chemical shift anisotropies (CSAs) are presented. In addition results for implementing these techniques on a fragment of the prion protein are presented, and their implications are discussed. A technique which provides sensitivity enhancement in multiple-quantum (MQ) NMR experiments is described, and promising results are shown for model systems. Finally, the extension of the sensitivity enhancement technique described previously is employed in a novel experiment to extract structural information. The results of this experiment on a model system are very encouraging.;Another realm in which advances discussed in this dissertation have been made is the field of oriented systems. The liquid crystal phase has been successfully employed to provide relevant structural information about small molecules co-dissolved within it. The methods by which this information was obtained rely on a unique switched angle spinning (SAS) probe design; the design and performance of this probe is discussed within this dissertation. By varying the spinning axis, interactions that provide relevant structural information are introduced to a controlled degree. Within the realm of oriented systems, work has been performed upon a unique oriented phase: bicelles. These unique oriented bilayered micelles provide an environment which mimics the native environment for membrane proteins. Work presented in this dissertation details current results and future efforts to aid in our understanding of the native structure of these vital proteins.