title: Structural alterations of human serum albumin caused by glycative and oxidative stressors revealed by circular dichroism analysis.
abstract: The aim of this work was to evaluate the ability of oxidative and glycative stressors to modify properties of human serum albumin (HSA) by analyzing markers of glycation (pentosidine) and oxidation (advanced oxidative protein products (AOPPs)) and assessing fluorescence and circular dichroism. HSA was incubated for up to 21 days with ribose, ascorbic acid (AA) and diethylenetriamine pentacetate (DTPA) in various combinations in order to evaluate influences of these substances on the structure of HSA. Ribose was included as a strong glycative molecule, AA as a modulator of oxidative stress, and DTPA as an inhibitor of metal-catalyzed oxidation. Ribose induced a significant increase in pentosidine levels. AA and DTPA prevented the accumulation of pentosidine, especially at later time points. Ribose induced a mild increase in AOPP formation, while AA was a strong inducer of AOPP formation. Ribose, in combination with AA, further increased the formation of AOPP. DTPA prevented the AA-induced generation of AOPP. Ribose was also a potent inducer of fluorescence at 335nm ex/385nm em, which is typical of pentosidine. AA and DTPA prevented this fluorescence. Circular dichroism showed complex results, in which AA and DTPA were strong modifiers of the percentages of the alpha-helical structure of HSA, while ribose affected the structure of HSA only at later time points.
authors.affiliation: Department of Internal Medicine, University of Genova, Genova 16132, Italy. nicola.traverso@unige.it.
authors.forename: Fiammetta
authors.initials: F
authors.lastname: Monacelli
authors.forename: Daniela
authors.initials: D
authors.lastname: Storace
authors.forename: Cristina
authors.initials: C
authors.lastname: D'Arrigo
authors.forename: Roberta
authors.initials: R
authors.lastname: Sanguineti
authors.forename: Roberta
authors.initials: R
authors.lastname: Borghi
authors.forename: Davide
authors.initials: D
authors.lastname: Pacini
authors.forename: Anna L
authors.initials: AL
authors.lastname: Furfaro
authors.forename: Maria A
authors.initials: MA
authors.lastname: Pronzato
authors.forename: Patrizio
authors.initials: P
authors.lastname: Odetti
authors.forename: Nicola
authors.initials: N
authors.lastname: Traverso
mesh_terms: D000085:Acetates / Q000494:pharmacology; D001120:Arginine / Q000031:analogs & derivatives / Q000494:pharmacology; D001205:Ascorbic Acid / Q000494:pharmacology; D002942:Circular Dichroism; D005453:Fluorescence; D006031:Glycosylation / Q000187:drug effects; D006801:Humans; D008239:Lysine / Q000031:analogs & derivatives / Q000494:pharmacology; D010084:Oxidation-Reduction / Q000187:drug effects; D018384:Oxidative Stress / Q000187:drug effects; D017433:Protein Structure, Secondary; D012709:Serum Albumin / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D000085:Acetates; D012709:Serum Albumin; D001120:Arginine; C062187:pentosidine; D008239:Lysine; D001205:Ascorbic Acid