Hen egg white lysozyme fibrillation: a deep-UV resonance Raman spectroscopic study.
Amyloid fibrils are associated with numerous degenerative diseases. The molecular mechanism of the structural transformation of native protein to the highly ordered cross-beta structure, the key feature of amyloid fibrils, is under active investigation. Conventional biophysical methods have limited application in addressing the problem because of the heterogeneous nature of the system. In this study, we demonstrated that deep-UV resonance Raman (DUVRR) spectroscopy in combination with circular dichroism (CD) and intrinsic tryptophan fluorescence allowed for quantitative characterization of protein structural evolution at all stages of hen egg white lysozyme fibrillation in vitro. DUVRR spectroscopy was found to be complimentary to the far-UV CD because it is (i) more sensitive to beta -sheet than to alpha -helix, and (ii) capable of characterizing quantitatively inhomogeneous and highly light-scattering samples. In addition, phenylalanine, a natural DUVRR spectroscopic biomarker of protein structural rearrangements, exhibited substantial changes in the Raman cross section of the 1000-cm(-1) band at various stages of fibrillation.
Xu
Ming
M
Department of Chemistry, University at Albany, SUNY, 1400 Washington Ave., Albany, NY 12222, USA.
Ermolenkov
Vladimir V
VV
Uversky
Vladimir N
VN
Lednev
Igor K
IK
D000682:Amyloid / Q000737:chemistry; D000818:Animals; D002645:Chickens; D002942:Circular Dichroism; D005260:Female; D006801:Humans; D066298:In Vitro Techniques; D018625:Microscopy, Atomic Force; D009113:Muramidase / Q000737:chemistry*; D055620:Optical Phenomena; D010063:Ovum / Q000201:enzymology; D010649:Phenylalanine / Q000737:chemistry; D011489:Protein Denaturation; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet / Q000379:methods; D013059:Spectrum Analysis, Raman / Q000379:methods*; D014364:Tryptophan / Q000737:chemistry
D000682:Amyloid; D010649:Phenylalanine; D014364:Tryptophan; D009113:Muramidase