| The use of Solid State Nuclear Magnetic Resonance (NMR) Spectroscopy for macromolecules was once precluded by low sensitivity, low resolution, lack of sequential assignment methods applicable to large molecules, and lack of methods for determining large sets of structural constraints in isotopically enriched molecules. Rapid progress has been made recently which clears many of these hurdles, and many proof-of-principle experiments which focus upon small model peptides have been reported. Presently many isotopic labelling schemes are being explored in a number of laboratories with the ambition of adapting experimental solid state NMR approaches to extract distance constraints for an entire protein.; This thesis concentrates on developing a means to gauge in advance the potential of a given labelling scheme to provide ample, accurate, precise, and structurally-meaningful data. We develop a framework for this analysis which focuses upon two of the most unavoidable sources of error, instrument noise and spin-diffusion, and assesses their impact on a straightforward magnetization-exchange solid state NMR experiment.; We begin by providing a qualitative assessment of the problem one faces in choosing solid state NMR experiments on fully labelled protein. We then develop one conceivable way to assess the quality of dipolar recoupling data from the given spin system by simulating data using Monte Carlo numerical methods, and employing an analytic transform of the data to calculate theoretical experimentally-derived internuclear distance-dependent rates, similar to the rationale already used by rate-matrix refinement procedures for Nuclear Overhauser Spectroscopy (NOESY) data. The analysis provides a method for comparing the relative merits of different solid state NMR approaches to the structure determination of large molecules. A framework is also developed for quantitatively segregating dipolar coupling constraints according to the quality of their information content, and limits on precision of measurement are suggested. |
