Fundamentals and Applications of Biological EPR Spectroscopy

Abstract

This thesis details robust experimental investigations of sample conditions for EPR spectroscopy. This work investigates two areas of sample conditions. First, a robust experimental investigation of the effect of sample matrix deuteration on relaxation behavior is presented. This work serves to guide experimental practitioners to choose optimal sample conditions for EPR measurement. Second, an investigation of the effects of different spin labels on intra-protein distance measurements in double electron electron resonance (DEER) spectroscopy is detailed. The distance distribution P(r) obtained via DEER includes the flexibility of the spin label in addition to the flexibility of the protein. This work provides a direct comparison of commonly used spin labels in DEER experiments on the same site pairs to isolate spin label effects on distribution broadening.