Spectroscopic Characterization of Amyloid Fibril Formation by Lysozyme

Hen egg white lysozyme is used as a model protein to study amyloid fibril formation (fibrillation). This three-week laboratory experience has been successfully implemented in an upper-level biochemistry course to introduce students to issues of protein conformation, spectroscopic characterization of conformational changes, and misfolding of proteins related to disease. The fluorescence and absorbance of amyloid-specific dyes are used to detect fibrillation; accompanying changes in secondary structure are revealed by circular dichroism. Potential small molecule fibrillation inhibitors are tested, and kinetics experiments are carried out to probe the effect of seeding with preformed fibrils on the rate of fibrillation.