Effect of mid-infrared free-electron laser irradiation on refolding of amyloid-like fibrils of lysozyme into native form.
Aggregation of lysozyme in an acidic solution generates inactive amyloid-like fibrils, with a broad infrared peak appearing at 1,610-1,630 cm(-1), characteristic of a β-sheet rich structure. We report here that spontaneous refolding of these fibrils in water could be promoted by mid-infrared free-electron laser (mid-IR FEL) irradiation targeting the amide bands. The Fourier transform infrared spectrum of the fibrils reflected a β-sheet content that was as low as that of the native structure, following FEL irradiation at 1,620 cm(-1) (amide I band); both transmission-electron microscopy imaging and Congo Red assay results also demonstrated a reduced fibril structure, and the enzymatic activity of lysozyme fibrils recovered to 70-90 % of the native form. Both irradiations at 1,535 cm(-1)(amide II band) and 1,240 cm(-1) (amide III band) were also more effective for the refolding of the fibrils than mere heating in the absence of FEL. On the contrary, either irradiation at 1,100 or 2,000 cm(-1) afforded only about 60 % recovery of lysozyme activity. These results indicate that the specific FEL irradiation tuned to amide bands is efficient in refolding of lysozyme fibrils into native form.
IR Free Electron Laser Research Center, Research Institute for Science and Technology (RIST), Tokyo University of Science, Yamazaki, Noda, Chiba, Japan. kawasaki@rs.tus.ac.jp
Takayasu
T
Kawasaki
Jun
J
Fujioka
Takayuki
T
Imai
Koichi
K
Tsukiyama
D000682:Amyloid / Q000737:chemistry / Q000378:metabolism / Q000528:radiation effects*; D000818:Animals; D002645:Chickens; D003224:Congo Red; D007259:Infrared Rays; D007834:Lasers; D009113:Muramidase / Q000737:chemistry / Q000378:metabolism / Q000528:radiation effects*; D011485:Protein Binding; D011487:Protein Conformation; D058849:Protein Refolding / Q000528:radiation effects*; D013057:Spectrum Analysis
D000682:Amyloid; D003224:Congo Red; C421864:hen egg lysozyme; D009113:Muramidase