title: Use of Raman spectroscopy for the identification of radical-mediated damages in human serum albumin.
abstract: Damages induced by free radicals on human serum albumin (HSA), the most prominent protein in plasma, were investigated by Raman spectroscopy. HSA underwent oxidative and reductive radical stress. Gamma-irradiation was used to simulate the endogenous formation of reactive radical species such as hydrogen atoms ((•)H), solvated electrons (e(aq)(-)) and hydroxyl radicals ((•)OH). Raman spectroscopy was shown to be a useful tool in identifying conformational changes of the protein structure and specific damages occurring at sensitive amino acid sites. In particular, the analysis of the S-S stretching region suggested the radical species caused modifications in the 17 disulphide bridges of HSA. The concomitant action of e(aq)(-) and (•)H atoms caused the formation of cyclic disulphide bridges, showing how cystine pairs act as efficient interceptors of reducing species, by direct scavenging and electron transfer reactions within the protein. This conclusion was further confirmed by the modifications visible in the Raman bands due to Phe and Tyr residues. As regards to protein folding, both oxidative and reductive radical stresses were able to cause a loss in α-helix content, although the latter remains the most abundant secondary structure component. β-turns motifs significantly increased as a consequence of the synergic action of e(aq)(-) and (•)H atoms, whereas a larger increase in the β-sheet content was found following the exposure to (•)OH and/or (•)H attack.
authors.affiliation: Istituto I.S.O.F., Consiglio Nazionale delle Ricerche, Bologna, Italy.
authors.forename: Z
authors.initials: Z
authors.lastname: Jurasekova
authors.forename: A
authors.initials: A
authors.lastname: Tinti
authors.forename: A
authors.initials: A
authors.lastname: Torreggiani
mesh_terms: D003553:Cystine / Q000737:chemistry; D004220:Disulfides / Q000737:chemistry; D005609:Free Radicals / Q000737:chemistry*; D006801:Humans; D008958:Models, Molecular; D010084:Oxidation-Reduction; D011487:Protein Conformation; D017510:Protein Folding; D012709:Serum Albumin / Q000737:chemistry*; D013059:Spectrum Analysis, Raman / Q000379:methods
chemical_list: D004220:Disulfides; D005609:Free Radicals; D012709:Serum Albumin; D003553:Cystine