Hen egg white lysozyme fibrillation: a deep‐UV resonance Raman spectroscopic study |
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| Authors: | Ming Xu Vladimir V Ermolenkov Vladimir N Uversky Igor K Lednev |
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| Institution: | 1. Department of Chemistry, University at Albany, SUNY, 1400 Washington Ave., Albany, NY 12222, USA;2. Center for Computational Biology and Bioinformatics, Department of Biochemistry and Molecular Biology, Indiana University School of Medicine, Indianapolis, 714 North Senate Ave., Suite 250, IN 46202, USA;3. Institute for Biological Instrumentation, Russian Academy of Sciences, Pushchino, Moscow Region 142292, Russia |
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| Abstract: | Amyloid fibrils are associated with numerous degenerative diseases. The molecular mechanism of the structural transformation of native protein to the highly ordered cross‐β 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 β ‐sheet than to α ‐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. (© 2008 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim) |
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| Keywords: | Hen egg white lysozyme deep‐UV resonance Raman spectroscopy tryptophan fluorescence spectroscopy circular dichroism spectroscopy amyloid fibril fibrillation |
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