Hydration structure analysis of lysozyme amyloid fibrils by thermally stimulated depolarization currents (TSDC) technique

Abstract

Thermally stimulated depolarization currents technique has been employed to investigate the conformation of hen egg white lysozyme in native and amyloid form, in the state of powder at very low hydration level. The technique, able to detect the current generated by thermally activated reorientation of water dipoles previously oriented by an electric field, exploits H2O dipoles, belonging to the solvation shell, as a probe to gain information on the protein conformation. Large differences are detected between the TSDC spectra related to the two different protein conformations, for what concerns the number and position of the main peaks, the native form displaying two peaks, at T M=175 K and at TM=297 K, and the amyloid one, only one at intermediate temperature (TM=235 K). The spectra have been compared with those monitored for poly-L-lysine (MW 80400), as received and prepared in different ways, i.e. α-helix, β-sheet, and coil conformation, respectively. The poly-L-lysine spectra show specific features that can be attributed to water texture around the secondary structure adopted by the macromolecule: the results stress how TSDC technique is a tool of great potential value in the conformational analysis of proteins. © 2008 - IOS Press and the authors. All rights reserved.