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doi:10.1016/j.molstruc.2004.11.076
Copyright © 2004 Elsevier B.V. All rights reserved.
Radiation damage of lysozyme in a biomimetic model: some insights by
Raman spectroscopy
Armida Torreggiani
a, ,
, Maurizio Tamba
a,
, Immacolata Manco
b,
, Maria Rosaria Faraone-Mennella
b,
, Carla Ferreri
a,
and Chryssostomos Chatgilialoglu
a,
a
Istituto ISOF, Consiglio Nazionale delle Ricerche, Via P. Gobetti 101, Bologna 40129, Italy
b
Dip. Chimica Biologica, Università di Napoli, via Mezzocannone 16, Napoli 80134, Italy
Received 7 September 2004; revised 18 November 2004; accepted 18 November 2004. Available online 11
January 2005.
Journal of Molecular Structure
Volumes 744-747, 3 June 2005, Pages 767-773
MOLECULAR SPECTROSCOPY AND MOLECULAR STRUCTURE 2004
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Abstract
A biomimetic model for protein/lipid radical damage is described. The model shows that the protein
degradation is accompanied by trans lipid isomerization due to thiyl radicals, derived from sulfur-
containing residues and diffused through the lipid bilayer. γ-Irradiation was used to obtain free
radical generation and lysozyme was chosen as enzyme example.
A detailed study of the Raman spectra of lysozyme irradiated at different doses coupled to vesicle
experiments, as well as enzymatic assays, is reported to gather a comprehensive view of the
irradiation effect.
The lysozyme resistance to degradation by γ-irradiation is remarkable and the protein structure play
a significant role in blocking the ready access of free radicals both to the sulfur-containing residues
and the active site. The degradation of sulfur moieties does not occur immediately, but as irradiation
progressed, the involvement of these residues takes place. In fact, structural changes induced on
the protein render these residues more exposed and susceptible of radical attack. Tyr are
confirmed to be among the most sensitive residues towards oxidation.
These results can be relevant for a better understanding of mechanisms of tandem radical
damages occurring in a biological environment.
Keywords: Lysozyme; γ-Irradiation; Raman spectroscopy; Radical damage; Trans lipid
Article Outline
1. Introduction
2. Experimental
2.1. Materials and methods
2.2. γ-Irradiation of DOPC-Lyso vesicles
2.3. γ-Irradiation of Lyso in aqueous solution
2.4. Activity assays
3. Results and discussion
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